JP3342013B2 - Dipeptidylquinazolones as anticancer agents - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a novel anticancer agent, and more particularly, the present invention relates to a quinazoline derivative having anticancer activity.

One group of anticancer drugs are antimetabolites with antifolate activity,
Examples include aminopterin and methotrexate. A relatively new compound of this type that has shown considerable promise in clinical trials is known as CB3717 and is described and claimed in GB 2 065 653B. However, despite its promising activity against human breast, uterine and liver cancer, CB3717 shows toxic symptoms in humans, especially to the liver and kidneys. These disadvantageous side effects are described in UK Patent No. 217, respectively.
5 903 and 2 188 319 as described and claimed in the absence of the 2-amino substituent of CB3717,
Or lower for compounds substituted with any of the various alternative substituents.

Compounds of this type are thought to act as anticancer agents by inhibiting the enzyme thymidylate synthase, which catalyzes the methylation of deoxyuridine monophosphate to produce the thymidine monophosphate required for DNA synthesis. The anti-cancer activity of CB3717 and similar compounds was determined by measuring their inhibitory effect on the enzyme in vitro and in cell cultures in cancer cell lines such as the mouse lymphoma cell line L1210 and the human breast cancer cell line MCF.
-7 can be assessed by measuring their inhibitory effect.

Antimetabolites, such as aminopterin and methotrexate, which are inhibitors of enzymes utilizing folate derivatives, also show promise in the treatment of various allergic diseases, such as allergic rhinitis, atopic dermatitis and psoriasis.

We now show that certain quinazoline derivatives not only show good levels of activity, especially with regard to their ability to inhibit thymidine synthase, but also exhibit a different mode of action than CB3717 and other related quinazoline derivatives reported. It was also found to show. For example, CB3717, especially UK Patent No. 2
Although the 2-methyl analog described and claimed in 188 319 produces antitumor activity through an intracellular polyglutamate form, the compounds of the present invention do not undergo a significant degree of γ-glutamylation. It is thought to act directly. This different mode of action of the compounds of the present invention offers the possibility of more tightly controlling the administration of these compounds to cancer patients. This is especially due to the shorter intracellular retention period after the end of administration and the lack of reliance on polyglutamylation, which can vary in degree from patient to patient. Furthermore, the replacement of the L-glutamic acid residue of CB3717 with another group in the compounds of the present invention confers different physical properties, which affects the overall properties of the compound.

Accordingly, the present invention comprises quinazolines of formula (I):
− Wherein R ¹ is hydrogen or amino; or R ¹ is alkyl, alkoxy or alkylthio, each having up to 6 carbon atoms; or R ¹ is aryl or aryl, each having up to 10 carbon atoms. Or R ¹ is halogeno, hydroxy or mercapto; or R ¹ is alkyl having up to 3 carbon atoms, halogeno, hydroxy and alkanoyl each having up to 6 carbon atoms. R ¹ is alkoxy having up to 3 carbon atoms, wherein R ¹ is alkoxy having up to 3 carbon atoms, and hydroxy and alkoxy having up to 6 carbon atoms. is intended to possess one or more substituents selected from; R ² is hydrogen or, Alkyl, alkenyl, alkynyl, hydroxyalkyl, alkoxyalkyl, mercaptoalkyl, alkylthioalkyl, halogenoalkyl, cyanoalkyl, aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl, alkanoylalkyl, carboxyalkyl, having up to 6 carbon atoms. Carbamoylalkyl or alkanoyl; Ar is phenylene or heterocyclene, which is unsubstituted or has halogeno, cyano, nitro, hydroxy, amino and carbamoyl, each having up to 6 carbon atoms alkyl, alkoxy, halogenoalkyl, possess one or more substituents selected from alkanoylamino and alkoxycarbonyl; R ³ is a dipeptide residue A is, it bound to the carbonyl group of COR ³ first 1N- terminal amino acid residue is an amino acid residue Wherein A is a carbon-carbon single bond or an alkylene group having up to 5 carbon atoms, V and V ′
Is each independently hydrogen or alkyl, alkenyl or alkynyl, each having up to 6 carbon atoms; the second amino acid residue is an α-amino acid residue;
Provided that one or more of the following conditions are met: (a) at least one of V and V 'is other than hydrogen; (b) the nitrogen atom of the second amino acid residue has at most 4 carbon atoms. An alkyl, alkenyl or alkynyl group, or an alkyl group having up to 3 carbon atoms carrying one or more halogeno or hydroxy substituents, or the α-amino acid residue of the second amino acid residue.
R ⁴ is hydrogen, or up to 4 carbon atoms, substituted with an ethylene, trimethylene or tetramethylene group attached to a carbon atom, and (c) the α-carbon atom of the second amino acid residue is completely substituted. R ⁵ is hydrogen, or alkyl having up to 4 carbon atoms; R ⁶ , R ⁷ and R ⁸ are each hydrogen, or alkyl having up to 4 carbon atoms or Alkoxy, or halogeno; these quinazolines may optionally be in the form of pharmaceutically acceptable salts, esters or amides.

A corresponding dipeptide having an L, L configuration, which is not one of the following: (a) being substituted at a carbon atom in the first amino acid residue adjacent to an amide bond between the two residues The present invention as compared to (b) those that are N-substituted at the second amino acid residue and (c) those that are completely substituted at the α-carbon atom of the second amino acid residue. The quinazoline dipeptides are resistant to cleavage of the central amide bond of the dipeptide in vivo. Thus, the presence of one or more of these three forms of substitution adjacent to the amide bond results in such cleavage resistance. Without being limited to any particular theory, in the quinazolines of the present invention, substitutions of the type present adjacent to an amide bond between two residues of a dipeptide may occur due to steric hindrance around the bond. It is expected to achieve the effect.

Although each of these forms of substitution may exist in the compound of the present invention, it is preferred that two or less, particularly only one, of these forms is present, and the substitution at the nitrogen atom of the second amino acid residue is extremely preferable. Thus, in a preferred aspect of the invention, R ³ is a dipeptide residue,
The first N-terminal amino acid residue attached to the carbonyl group of COR ³ is an amino acid residue Wherein A is a carbon-carbon single bond or an alkylene group having up to 5 carbon atoms, V and V ′
Is each independently hydrogen, or alkyl, alkenyl or alkynyl, each having up to 6 carbon atoms; the second amino acid residue is an α-amino acid residue and has up to 4 carbon atoms at the nitrogen atom. Having an alkyl, alkenyl or alkynyl group, or 1
An ethylene, trimethylene or tetramethylene group bound to an alkyl group having up to 3 carbon atoms or carrying an α-carbon atom of a second amino acid residue carrying one or more halogeno or hydroxy substituents , For example, α-amino acid residues described below.

As used herein, alkyl, alkenyl, alkynyl and alkylene include both straight-chain and branched-chain groups, but individual alkyl or alkylene groups such as "propyl" or "propylene" are specified only as straight-chain groups. . Similar arrangements apply to other generic terms. Further, the numbering system used for the quinazoline nucleus is conventional as shown below.

Amino acid residues are tabulated using standard methods (Pure and Appli
ed Chemistry, 1974, 40 , 317 and European Journal of
Biochemistry, 1984, 138 , 9). However, to avoid uncertainty, γ-glutamyl is optionally substituted with the group H ₂ NCH (CO 2
It should be noted that ₂ H) CH ₂ CH ₂ CO— or —HNCH (CO ₂ H) CH ₂ CH ₂ CO— means that in these groups the carbon atom is one carbon atom of the α-carbonyl group. These numbers are assigned as places.

Quinazolines of the present invention (present in the peptide residue R ³⁾ comprises at least one asymmetric carbon atom, thus it may be present as an optically active form it will be appreciated. It is to be understood that the present invention also encompasses various optically active forms of quinazoline, and it is common knowledge to obtain these optically active forms by stereospecific synthesis or by separating mixtures of isomeric compounds. However, one isomer may be more important than the other due to the nature of the activity it exhibits or due to its outstanding physical properties, such as water solubility.

Where R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ or R ⁸ are alkyl, or any of which are suitable for an alkyl substituent in Ar, for example, methyl, ethyl, propyl Or isopropyl.

Suitable when R ² is alkenyl are, for example, prop-2-enyl, buta-
2-enyl, but-3-enyl, 2-methylprop-2
-Enyl, hexa-2-enyl, hexa-5-enyl,
Or 2,3-dimethylbut-2-enyl.

Suitable when R ² is alkynyl are, for example, prop-2-ynyl, buta-
2-ynyl, but-3-ynyl, penta-2-ynyl,
3-methylpenta-4-ynyl, hexa-2-ynyl,
Or hexa-5-ynyl.

A suitable value for any of R ¹ , R ⁶ , R ⁷ or R ⁸ when it is alkoxy, or an alkoxy substituent in Ar, is, for example, methoxy, ethoxy or isopropoxy.

Where R ¹ is alkylthio, suitable are
For example, methylthio or isopropylthio.

Suitable when R ¹ is aryl is, for example, phenyl or tolyl.

Suitable when R ¹ is aryloxy is, for example, phenoxy or tolyloxy.

Suitable when R ¹ , R ⁶ , R ⁷ or RR ⁸ is halogeno are, for example, fluoro, chloro, bromo or iodo.

Suitable when R ¹ is substituted alkyl is
For example, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 3-fluoropropyl, chloromethyl, dichloromethyl, hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl,
Acetamidomethyl, 3-acetamidopropyl, or propionamidomethyl.

Suitable when R ¹ is substituted alkoxy are, for example, 2-hydroxyethoxy, 4-hydroxybutoxy, 3-hydroxy-2-methylpropoxy,
-Methoxyethoxy, 3-methoxypropoxy, or 2-ethoxyethoxy.

Suitable when R ² is hydroxyalkyl, alkoxyalkyl, mercaptoalkyl or alkylthioalkyl are, for example, 2-hydroxyethyl, 3-hydroxypropyl, 2-methoxyethyl,
-Ethoxyethyl, 3-methoxypropyl, 2-methoxypropyl, 2-mercaptoethyl, 3-mercaptopropyl, 2-methylthioethyl, 3-methylthiopropyl or 2-ethylthioethyl.

Suitable when R ² is halogenoalkyl, cyanoalkyl, aminoalkyl, alkylaminoalkyl or dialkylaminoalkyl are, for example, 2-fluoroethyl, 2-chloroethyl, 2-bromoethyl, 3-fluoropropyl, 3 -Chloropropyl, cyanomethyl, 2-cyanoethyl, 3-cyanopropyl,
2-aminoethyl, 3-aminopropyl, 3-amino-
2-methylpropyl, 2-methylaminoethyl, 2-dimethylaminoethyl, 2-ethylaminoethyl, 2-diethylaminoethyl, 3-methylaminopropyl, or 3-dimethylaminopropyl.

Suitable when R ² is alkanoylalkyl, carboxyalkyl, carbamoylalkyl or alkanoyl are, for example, acetonyl, 2-acetylethyl, propionylmethyl, 2-propionylethyl, 3-acetylpropyl, 4-acetylbutyl, Carboxymethyl, 2-carboxyethyl, carbamoylmethyl, acetyl, propionyl, or butyryl.

Suitable when Ar is phenylene are, for example, 1,3- or especially 1,4-phenylene.

Suitable when Ar is heterocyclene are, for example, 5- or 6-membered aromatic (ie fully unsaturated) heterocyclene-diradicals, such as oxygen,
Those containing up to two heteroatoms selected from the group consisting of nitrogen and sulfur, such as thienylene, pyridylene, pyrimidinylene, thiazolylen, or oxazolylen. Of particular interest are compounds where Ar is pyrimidinylene, especially pyridylene or especially phenylene.

Suitable halogeno, halogenoalkyl, alkanoylamino, or alkoxycarbonyl substituents in Ar are, for example, fluoro, chloro, bromo, iodo, fluoromethyl, difluoromethyl, trifluoromethyl, acetamido, propionamido, isopropionamido, methoxycarbonyl , Ethoxycarbonyl, or isobutoxycarbonyl.

When substituents are present in Ar, a suitable number of substituents is three substituents, in particular two substituents, or especially one substituent; and one or two substituents are Conveniently adjacent to the atom attached to COR ³ , a halogeno substituent, for example fluoro, is preferred.

The first amino acid residue of R ³ is an α-amino acid, and the second amino acid residue may be an α-amino acid, wherein α-amino acid
The carbon atoms are asymmetric, that is, the carbon atoms are bonded to four different groups or atoms, one of which is a carboxy group, and the other is a group bonded to that carbon atom. N <. If the second amino acid residue contains a fully substituted α-carbon atom, none of the four bonds to that α-carbon atom will attach it to a hydrogen atom, provided that two of the substituents May be the same,
Thus, the α-carbon atom need not be asymmetric.

The first amino acid residue of R ³ of the peptide residue is the asymmetric α
The carbon atom may be in either the L- or D- configuration, and the second amino acid residue may be in any configuration if it is asymmetric at its α-carbon atom. However, it is preferred that the first amino acid residue and, where appropriate, also the second amino acid residue are of the L-configuration. V
And both V 'are other than hydrogen and are different groups, the carbon atoms to which they are attached are also asymmetric, resulting in erythro and threo forms for the amino acid forming the first residue. It will be apparent that they may both exist as optically active isomers.

While the compounds of the present invention may exist as a mixture of stereoisomers, it is preferable to resolve them into one optically active isomer. Since the synthesis of such required compounds is complicated, it is preferable that they contain as few asymmetric carbon atoms as possible, as long as they achieve the desired activity. Thus, for example, if one of V and V 'is other than hydrogen, then if both V and V' are other than hydrogen and are the same group, the presence of an additional asymmetric carbon atom Will be avoided. But often,
Said to in R ³ as there are two asymmetric carbon atoms, preferably L, L-quinazoline dipeptide is advantageous to substantially free of D, D-dipeptide, and it L, D
Advantageously, they are also substantially free of-and D, L-dipeptides. Thus, in its preferred form, the dipeptide is substantially free of all three other isomers. The term "substantially free" is used herein to indicate that the presence of the other said isomer does not exceed 20% by weight, in particular does not exceed 10% by weight.

Suitable for R ³ is, first 1N- terminal amino acid residue Is the second amino acid residue Combined with the expression Of a dipeptide residue giving group R ^3, A in these formulas, V and V 'are those previously defined; X is alkyl having hydrogen, up to 4 carbon atoms, alkenyl or alkynyl, Or a C _1-3 alkyl group (substituted with one or more halogeno substituents or hydroxy groups); Y is hydrogen or alkyl, alkenyl or alkynyl, each having up to 6 carbon atoms. Or X and Y together are a group (CH ₂ ) _n, where n is 2, 3 or 4; Y ′ is hydrogen or alkyl having up to 6 carbon atoms each , Alkenyl or alkynyl; or Y 'is alkyl having up to 6 carbon atoms and is selected from amino, carboxy, hydroxy and mercapto. Or carries one or more substituents; or Y 'is phenyl or benzyl; provided one or more of the following: (a) at least one of V and V' is hydrogen (B) X is other than hydrogen; and (c) Y and Y 'are each other than hydrogen.

Another suitable for R ³ is, first 1N- terminal amino acid residue (A, V and V 'are as defined above) is, the type of -N (X) C (Y) (Y') - non-CO ₂ H,
Residues of natural amino acids, or alkyl, alkenyl or alkynyl having up to 4 carbon atoms, or C
A dipeptide residue linked to a second amino acid residue corresponding to an amino acid residue N-substituted by an _1-3 alkyl group (substituted by one or more halogeno groups or hydroxy groups) .

Suitable for A are a carbon-carbon single bond or, preferably, an alkylene group of 2 carbon atoms, or especially of 1 carbon atom, for example CH ₂ CH ₂ , especially CH ₂ .

Which V and V 'are suitable for it when it is alkyl, alkenyl or alkynyl is, for example those described above with respect to groups such as R ^2, in particular methyl.

When V is other than hydrogen, V 'is also preferably other than hydrogen, and is conveniently the same as V. One group of compounds of particular interest is the group A-CV
Another group of compounds of minor interest, including (V '), where V and V' are each hydrogen, is the group A-
CV (V '), where V and V' are each the same alkyl group, especially a methyl group. Thus many atomic group A-CV in the compound (V ') is a group A', i.e. up to 6, eg 1, 3, or particularly an alkylene group having 2 carbon atoms, for example CH _2, CH ₂ CH ₂ C
H ₂ , especially CH ₂ CH ₂ , or especially CH ₂ CH (CH ₃ ), especially CH ₂ C if the nitrogen atom of the second amino acid residue is unsubstituted
Advantageously, (CH ₃ ) ₂ .

Alkyl X has up to 4 carbon atoms, suitable for it when it is alkenyl or alkynyl is, for example those described above with respect to groups such as R ^2, in particular prop-2-enyl, prop-2-ynyl , Especially C _1-3
An alkyl group is for example ethyl, especially methyl. Suitable when X is a halogeno-substituted or hydroxy-substituted C _1-3 alkyl group are those described above for groups such as R ¹ , for example, preferred halogeno substituents are fluoro groups, 2 for hydroxy substitution
-Substituted ethyl groups are particularly important, in the case of halogeno substitution substituted ethyl, especially methyl groups, are particularly important.
X is preferably a group other than hydrogen, which is conveniently any of the monovalent aliphatic hydrocarbon groups described above, or to a lesser extent together with Y together with the group (CH ₂ ) _n, where n is 2, 4, especially 3.

Suitable when Y and Y 'are alkyl are, for example, those described above for groups such as R ¹ ,
For example, propyl and isopropyl, ethyl, especially methyl.

That Y and Y 'are suitable therefor in the case of alkenyl or alkynyl, even those described above with respect to groups such as R ^2, in particular prop-2-enyl, and prop-2-ynyl.

Suitable when Y 'is a substituted alkyl group are those groups which carry only one substituent, in particular a carboxy group, which is conveniently on the terminal carbon atom of the alkyl group. is there. Of the various possibilities for Y ', these groups are particularly important. Of particular interest are alkyl groups having up to three carbon atoms, ie, methyl, ethyl, propyl, and isopropyl,
Larger groups can also be important, especially if they are branched. For example, this type of preferred groups Y 'is, CH ₂ CO ₂ H or _{CH 2 CH 2 CH 2 CO 2} H,, in particular CH ₂ CH (C
H ₃₎ CO ₂ H or _{CH (CH 3,) CH 2} CO 2 H, in particular CH ₂ CH ₂ CO ₂ H.

Natural amino acids H ₂ NCH (Y ′) CO ₂ H containing the group Y ′, which are present in the group R ³ of the quinazolines according to the invention as such or in an N-substituted, for example N-methyl-substituted form. (These groups Y ′ are the groups detailed above,
Or other forms of groups) are alanine (Y '= CH
₃ ), arginine (Y ′ = (CH ₂ ) ₃ NHC (NH ₂ ) = NH),
Aspartic acid (Y ′ = CH ₂ CO ₂ H), cysteine (Y ′
= CH ₂ SH), glutamic acid _{(Y '= CH 2 CH 2} CO 2 H), isoleucine _{(Y' = CH (CH 3} ) CH 2 CH 3), leucine (Y '
= CH ₂ CH (CH ₃ ) CH ₃ ), ornithine (Y ′ = (CH ₂ ) ₃ N
H _2), phenylalanine _{(Y '= CH 2 C 6} H 5), serine _{(Y' = CH 2 OH)} , and valine _{(Y '= CH (CH 3} ) 2)
It is.

The unnatural amino acid H ₂ NCH (Y ′) CO ₂ H containing the group Y ′, which is present as such or in the N-substituted, for example N-methyl-substituted form, in the group R ³ of the quinazolines of the invention examples of and can) is norvaline _{(Y '= CH 2 CH 2} CH 3), norleucine _{(Y' = (CH 2)} 3 CH 3), 2- phenylglycine _{(Y '= C 6 H 5} ), and tert -Leucine (Y '= C
(CH ₃ ) ₃ ).

Highly preferred groups R ³ was N- substituted includes a second amino acid residues in particular N- methyl-substituted. Thus, these groups R ³ have the formula Wherein A, V, V ', X, Y and Y' are as defined above, provided that X is other than hydrogen, especially methyl, preferably Y is hydrogen. Yes,
Advantageously, V and V 'are also hydrogen. Alternatively, the less preferred group R ³ is V ³
And at least one, and preferably both, of the first amino acid residue are other than hydrogen, or α-
It includes a second amino acid residue in which the carbon atoms have been completely substituted, in each case one of these substituents is preferably methyl, and both of these possibilities are optional, but preferred. It is preferably present to a lesser extent. Thus, these groups R ³ have the formula In which A, V, V'X, Y and Y 'are as defined above, provided that V and V' are each other than hydrogen, for example each methyl, and / Or Y and Y 'are each other than hydrogen, for example each methyl, and preferably X
Is hydrogen.

Particularly useful radicals R ³ has the formula _{-NHCH (CO 2 H) -A'-} CON (X) CH (Y ') - CO has ₂ H, A in the formula' has up to 6 carbon atoms Alkylene groups such as CH ₂ CH (CH ₃ ), CH ₂ C (CH ₃ ) ₂ , CH ₂
CH ₂ CH ₂ or CH ₂ CH ₂ , X is as defined above and Y ′ is methyl, ethyl, propyl, hydroxymethyl, phenyl or benzyl, especially of the formula —NHCH (CO ₂ H _{) -A'-CON (CH 3)} CH (CO 2 H) (CH 2) have _m -CO ₂ H, as a in the formula 'is as defined in the above, for example, CH ₂ C
_{H (CH 3), CH 2} C (CH 3) 2, in particular CH ₂ CH ₂ CH _2, in particular CH ₂ CH ₂
And m is 1, in particular 3, in particular 2.

A specific example of R ³ is the dipeptide γ-glutamyl-
α-methylalanine, γ-4-methylglutamylglutamic acid, γ-4,4-dimethylglutamylglutamic acid,
γ-glutamylproline, γ-glutamyl-N-methylglycine, γ-glutamyl-N-methylalanine, γ-
Glutamyl-N-methylaspartic acid, especially γ-glutamyl-N-methyl-2-aminoadipic acid, especially γ-
Glutamyl-N-methylglutamic acid, preferably their L, L form, or in the case of γ-glutamyl-N-methylglycine, the L form.

Suitable pharmaceutically acceptable salt forms of the quinazolines of the present invention include, for example, acid addition salts with inorganic or organic acids such as hydrochloric acid, hydrobromic acid, trifluoroacetic acid or maleic acid; or alkali metal salts , For example, sodium salts, alkaline earth metal salts, for example, calcium salts,
Or an ammonium salt such as a tetra (2-hydroxyethyl) ammonium salt.

Suitable pharmaceutically acceptable ester forms of the quinazolines of the invention are, for example, esters with aliphatic alcohols having up to 6 carbon atoms, for example methyl, ethyl or tert-butyl esters.

Form of quinazolines and pharmaceutically acceptable and can appropriate amino of the present invention, such as unsubstituted amides of formula -CONH _2,
Or especially benzyl substituted amide of the formula -CONHCH ₂ C ₆ H _5.

It should be understood that R ³ can include several carboxy groups. If it comprises three carboxy groups such as various preferred examples of dipeptide residues R ^3, e.g. R ³
Consists of two glutamic acid residues, the salt or ester is a monoacid-disalt or -ester or -amide, or a diacid-monosalt or -ester or -amide, or even a trisalt or -ester. Or it may be -amide.

Particularly preferred for the various symbols R ¹ -R ⁸ and Ar are indicated for the preferred quinazolines, respectively, below. Furthermore, in the case of R ⁴ , R ⁵ , R ⁶ and R ⁸ , compounds in which each is hydrogen are of particular interest. However, for R ^7, this is other than hydrogen, for example methoxy, any one of fluoro and chloro groups, especially alkyl groups, compounds such as methyl are also particularly important.

Preferred quinazolines of the invention are those having the above formula: wherein R ¹ is halogeno- or hydroxy-substituted alkyl, or especially hydrogen, amino, alkyl or alkoxy, especially fluoromethyl, hydroxymethyl, hydrogen, amino, methyl, ethyl or methoxy,; R ² is methyl, ethyl, prop-2-enyl, prop-2-ynyl, 2-hydroxyethyl, 3-hydroxypropyl, be a 2-fluoroethyl or acetonyl, Ar is 1,4-phenylene, 2-fluoro-1,4-phenylene or 2,6-difluoro-1,4-phenylene; R ³ is the dipeptide γ-glutamyl-N-methyl-2-
Aminoadipic acid, or γ-glutamyl-N-methylglutamic acid, preferably in the L, L form; R ⁴ is hydrogen or methyl; R ⁵ is hydrogen; R ⁶ is hydrogen or chloro R ⁷ is hydrogen, methyl, fluoro or chloro; and R ⁸ is hydrogen, methoxy or chloro.

Particularly preferred quinazolines of the invention are those having the above formula: wherein R ¹ is methyl; R ² is methyl, ethyl, or preferably prop-2-
Ar is 1,4-phenylene or 2-fluoro-1,4-phenylene; R ³ is γ-L-glutamyl-N-methyl-L-glutamic acid; R ⁴ is hydrogen or methyl by and; R ⁵ is hydrogen; R ⁶ is hydrogen or chloro; R ⁷ is hydrogen, methyl, methoxy, fluoro or chloro; and R ⁸ is hydrogen, methyl, methoxy or chloro.

Other quinazolines of particular importance according to the invention are R ¹ ,
R ² , R ⁴ —R ⁸ , and Ar have the same meaning, provided that R ³ has any of the above meanings. However, among the particularly preferred quinazolines of the present invention are: Np- [N- (3,4-dihydro-2-amino-4-
Oxoquinazolin-6-ylmethyl) -N-ethylamino] benzoyl-L-γ-glutamyl-N-methyl-L
-Glutamic acid, Np- [N- (3,4-dihydro-2-amino-4-
Oxoquinazolin-6-ylmethyl) -N-methylamino] benzoyl-L-γ-glutamyl-N-methyl-L
-Glutamic acid, Np- [N- (3,4-dihydro-2-amino-4-
Oxoquinazolin-6-ylmethyl) -N- (prop-
2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-methyl-4-
Oxoquinazolin-6-ylmethyl) -N-methylamino] benzoyl-L-γ-glutamyl-N-methyl-L
-Glutamic acid, Np- [N- (3,4-dihydro-2-methyl-4-
Oxoquinazolin-6-ylmethyl) -N-ethylamino] benzoyl-L-γ-glutamyl-N-methyl-L
-Glutamic acid, Np- [N- (3,4-dihydro-2-methyl-4-
Oxoquinazolin-6-ylmethyl) -N- (prop-
2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2,7-dimethyl-4)
-Oxoquinazolin-6-ylmethyl) -N-methylamino] benzoyl-L-γ-glutamyl-N-methyl-
L-glutamic acid, Np- [N- (3,4-dihydro-2,7-dimethyl-4
-Oxoquinazolin-6-ylmethyl) -N-ethylamino] benzoyl-L-γ-glutamyl-N-methyl-
L-glutamic acid, Np- [N- (3,4-dihydro-2,7-dimethyl-4
-Oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro- 2-methoxy-4
-Oxoquinazolin-6-ylmethyl) -N-methylamino] benzoyl-L-γ-glutamyl-N-methyl-
L-glutamic acid, Np- [N- (3,4-dihydro-2-methoxy-4)
-Oxoquinazolin-6-ylmethyl) -N-ethylamino] benzoyl-L-γ-glutamyl-N-methyl-
L-glutamic acid, Np- [N- (3,4-dihydro-2-methoxy-4)
-Oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro- 2-amino-4-
Oxoquinazolin-6-ylmethyl) -N-methylamino] -o-fluorobenzoyl-L-γ-glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-amino-4-
Oxoquinazolin-6-ylmethyl) -N-ethylamino] -o-fluorobenzoyl-L-γ-glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-amino-4-
Oxoquinazolin-6-ylmethyl) -N- (prop-
2-ynyl) amino] -o-fluorobenzoyl-L-
γ-glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-methyl-4-
Oxoquinazolin-6-ylmethyl) -N-methylamino] -o-fluorobenzoyl-L-γ-glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-methyl-4-
Oxoquinazolin-6-ylmethyl) -N-ethylamino] -o-fluorobenzoyl-L-γ-glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-methyl-4-
Oxoquinazolin-6-ylmethyl) -N- (prop-
2-ynyl) amino] -o-fluorobenzoyl-L-
γ-glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2,7-dimethyl-4
-Oxoquinazolin-6-ylmethyl) -N-methylamino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2, 7-dimethyl-4
-Oxoquinazolin-6-ylmethyl) -N-ethylamino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2, 7-dimethyl-4
-Oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] -o-fluorobenzoyl-L
-Γ-glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-methoxy-4
-Oxoquinazolin-6-ylmethyl) -N-methylamino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2- Methoxy-4
-Oxoquinazolin-6-ylmethyl) -N-ethylamino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid and Np- [N- (3,4-dihydro-2) -Methoxy-4
-Oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] -o-fluorobenzoyl-L
-Γ-glutamyl-N-methyl-L-glutamic acid, and their pharmaceutically acceptable salts, esters and amides.

The quinazolines of the present invention can be prepared by any method known to be applicable to the preparation of chemically related compounds.

A particularly preferred method for producing the quinazolines of the present invention is an acid of the formula: Or reacting their reactive derivatives with the terminal amino group of the dipeptide of the formula R ³ -H [R ¹ , R ² , R ³ ,
R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and Ar have the above-mentioned meanings, and R ¹ ,
The mercapto, amino and alkylamino groups in R ² , R ³ and Ar, and the carboxy group in R ¹ , R ² and Ar are all protected with common protecting groups, and R ¹ , R ² , R ^Three
And any of the hydroxy groups in Ar and the carboxy group in R ³ may be protected by common protecting groups, or those hydroxy or carboxy groups need not be protected; and R ⁹ is hydrogen or a protecting group], then including protecting group R ^9, and removing the non-target protecting group in a conventional manner.

In another method described in this method and the following compound R ³ -H, and as appropriate quinazoline acid, in asymmetric carbon atoms, conveniently to have the stereochemical configuration desired in the final quinazolines of formula (I) It is. Wherein on the protection of the amino group in R ³ is not of course apply to the amino group of the dipeptide R ³ -H.

Suitable reactive derivatives of the acids of the above formula are, for example, the following: acyl halides, for example acyl chlorides formed by the reaction of the above-mentioned acids with inorganic acid chlorides, for example thionyl chloride; Anhydrides formed by the reaction of an acid of formula I with a chloroformate such as isobutyl chloroformate; active esters,
For example, the above-mentioned acids and phenols such as pentafluorophenol, or alcohols such as 1-
Esters formed by reaction with hydroxybenzotriazole; reaction products of the above acids with carbodiimides, such as dicyclohexaylcarbodiimide; or especially acyl azides, such as those formed by reaction of the above acids with azides, such as diphenylphosphoryl azide. Azide, or an acyl phosphonate, such as an acyl formed by the reaction of an acid described above with a phosphonate, such as diethyl cyanophosphonate or (1H-1,2, -benzotriazol-1-yloxy) -tris (pyrrolidino) phosphonium hexafluorophosphate. Phosphonate.

A suitable protecting group for a hydroxy group is, for example, an ester-forming group, for example an acetyl or benzoyl group, which can be removed by hydrolysis with a base, for example sodium hydroxide, or wherein R ² is alkenyl or If it does not contain an alkynyl group, the protecting group may be, for example, an α-arylalkyl group, for example a benzyl group, which may be removed by hydrogenolysis over a catalyst, for example palladium.

A suitable protecting group for an amino group may be, for example, an alkoxycarbonyl group, for example a tert-butyloxycarbonyl group, which may be removed by treatment with an organic acid, for example trifluoroacetic acid; It may be an oxycarbonyl group or a trityl group, which can be removed by catalytic hydrogenation.

Other suitable protecting groups for primary amino groups are, for example, phthaloyl groups, which can be removed by treatment with an alkylamine, such as dimethylaminopropylamine, or with hydrazine.

A suitable protecting group for a carboxy group may be an ester-forming group, for example a tert-butyl group, which may be removed by treatment with an organic acid, for example trifluoroacetic acid. This avoids the possibility of racemisation that may occur for groups that can be removed with a base. However, it is sometimes appropriate to use a methyl group which can be removed by treatment with a base, for example sodium hydroxide.

Other suitable protecting groups for carboxy groups are, for example, allyl groups, which can be removed with a catalytic amount of tetrakistriphenylphosphine palladium (0) in the presence of an excess of pyrrolidine.

A suitable protecting group for a mercapto group is, for example, an ester-forming group, for example an acetyl group, which may be removed by hydrolysis with a base, for example sodium hydroxide.

While R ⁹ is preferably towards a hydrogen rather than a protecting group, those suitable for it when R ⁹ is a protecting group, for example, the pivaloyloxymethyl group. This group is a base,
Care can be taken to avoid racemisation, although it can be removed by hydrolysis, for example with sodium hydroxide.

Suitable protecting groups for various carboxy groups in R ³ are
Unintended protecting groups and protecting groups in R ¹ , R ² , R ³ and Ar
The product after removal of R ⁹ may be an ester-forming group that is included in the definition of quinazolines of the present invention. In these cases, esterified carboxy group in R ³ may be held in desired end product. Or it can also be used with different protecting groups in R ^3, which will be removed.

Dipeptide compound of the formula R ³ -H can also be obtained by any of a variety of common peptide synthesis methods described in the literature of peptide chemistry. Both classical methods involving reactions in solution or solid phase methods can be employed. However preferred peptide R ³ -H is prepared by reacting an appropriate 2 amino acids in solution, the amino group of the amino acid is R ³ -
Providing the N-terminal residue of H, wherein the carboxy group of the amino acid is
Provides C- terminal residues of R ³ -H, for example protected by the protective groups, in particular the respective suitable groups benzyloxycarbonyl or trityl group, and tert- butyl or allyl ester forming groups. Amino protecting group of the dipeptide will be necessarily removed before reaction with R ³ -H and quinazoline carboxylic acids, but existing carboxy protecting groups in the dipeptide would be advantageous to retain.

The carboxylic acid used as a starting material is a compound of the following formula: [Wherein R ¹ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ⁹ have the above-mentioned meaning, and Z is a replaceable group] by a compound of the following formula: —HNR ^{2 _{-Ar-CO 2 R 10 [R}} 2 and Ar in the formulas have the meanings given above, R ¹⁰ is a protecting group which can give a carboxylic acid by removing] is obtained by reacting with.

Z is, for example, a halogeno or sulfonyloxy group,
For example, a chloro, bromo, methanesulfonyloxy or toluene-p-sulfonyloxy group.

R ¹⁰ is, for example, a methyl or ethyl group, which can be removed by hydrolysis with a base, such as sodium hydroxide, or R ¹⁰ is, for example, t
It may be an ert-butyl group, which may be removed by cleavage with an organic acid such as trifluoroacetic acid.

The protecting group for the carboxy group in R ¹⁰ may be, for example, an ester-forming group, which retains the protecting groups for the mercapto, amino, carboxy and hydroxy groups in R ¹ , R ² and Ar It can be removed in a state.

An alternative method for producing the starting carboxylic acid is
This involves removing the residue from compounds of formula (I) where R ³ is instead an L-glutamic acid residue using the carboxypeptidase G2 enzyme.

In the quinazolines of the present invention, R ¹ is alkoxy, aryloxy, or alkoxy (up to 3) bearing one or more substituents (selected from hydroxy and alkoxy having up to 6 carbon atoms). Another preferred method for preparing those which have a carbon atom of is a compound of the formula: Is a compound of the formula: —HNR ² —Ar—COR ^{3 where} R ¹ has the last mentioned meaning;
² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , Ar and Z have the meanings given above, provided that R ² , R ³ and mercapto, amino, alkylamino and carboxy groups in Ar Is protected, for example, by the general protecting groups described above, and R ¹ , R ² , R ³ and
The hydroxy group in Ar may be protected, for example, with the general protecting groups described above, or the hydroxy group need not be protected]; And the cleavage of the R ¹ group at position 4 of the quinazoline ring by hydrolysis with a base, such as sodium hydroxide, to form the quinazolines of the present invention.

Another preferred method for preparing the quinazolines of the present invention wherein R ¹ is mercapto or alkylthio is a quinazoline of the following formula: Wherein R ¹ is halogeno or halogenoalkyl;
And R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and Ar have the above-mentioned meanings, provided that R ² , R ³ and mercapto, amino, alkylamino in Ar , Carboxy, and hydroxy groups may be protected, for example, with the general protecting groups described above, or amino, alkylamino, carboxy,
And the hydroxy groups need not be protected] with thiourea to give compounds where R ¹ is mercapto; or with alkylthiols to give compounds where R ¹ is alkylthio, arylthio; then it comprises a protecting group untargeted, including protecting group R ⁹ is removed by, for example, the conventional method.

Another preferred method for preparing the quinazolines of the present invention wherein R ¹ is alkylthio is a quinazoline of the formula: [Wherein R ¹ is mercapto and R ² , R ³ , R ⁴ , R ⁵ ,
R ⁶ , R ⁷ , R ⁸ , R ⁹ and Ar have the meaning described above, provided that
R ^2, R ³ and mercapto in Ar, amino, alkylamino, carboxy, and hydroxy group may be protected, for example the general protecting group, or an amino, alkylamino, carboxy, and hydroxy groups base need not] which is protected, for example, is reacted with ammonium hydroxide; then the resulting thiolate salt alkyl halide, for example by alkylation with methyl iodide; then protection of non-target, including protecting group R ⁹ Removing the group, for example, by the general methods described above.

An alternative method for preparing the quinazolines of the present invention is a compound of the formula: Is a compound of the formula: —HNR ² —Ar—COR ³ [wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹
Or, Ar has the above meaning, provided that R ¹ , R ³ and Ar
If there is a hydroxy group in, when there is a hydroxyalkyl group in R ¹ or R ^2, when there is a hydroxyalkoxy group in R ^1, when there is an amino group in R ^1, R ³ or Ar, R ² If there is an aminoalkyl group in, when there is an alkylaminoalkyl group in R ^2, when there is a carboxy or carboxyalkyl group in R ² or R ^3,
When there is a mercapto or mercaptoalkyl group in R ¹ , R ² or R ³ , the amino, carboxy and mercapto groups are protected, for example, by the above-mentioned general protecting groups, and the hydroxy group is, for example, the above-mentioned general protecting groups. Or the hydroxy group need not be protected; Z is a displaceable group]; and undesired protecting groups, including protecting group R ^9, can be Removal by a general method.

If a pharmaceutically acceptable salt of the novel compound of formula (I) is required, it can be obtained, for example, by reacting the compound with a suitable acid or base in a conventional manner.
If a pharmaceutically acceptable ester of the novel compound of formula (I) is required, it can be obtained, for example, by reacting the compound with a suitable acid or alcohol in a conventional manner. If a pharmaceutically acceptable amide of the novel compound of formula (I) is required, it can be obtained, for example, by reacting the compound or a suitable derivative thereof, such as acid chloride, with ammonia or a suitable amine. If an optically active form of the compound of formula (I) is required,
This can be obtained by performing any of the above methods using the optically active starting materials, or by resolving the racemic form of the compound in a conventional manner.

As mentioned above, quinazolines are at least partially due to their ability to inhibit the enzyme thymidylate synthase.
It is thought to function as an anticancer agent. This anti-drug activity
For example, it can be assessed by one or more of the following methods:-(a) In vitro assays that measure the ability of a test compound to inhibit the enzyme thymidylate synthase. Thymidylate synthase is obtained in partially purified form from L1210 mouse leukemia cells and is described in Jackman et al.
, 1986, 46 , 2810); (b) the test compound is used in cell culture in the leukemia cell line L121.
An assay that measures the ability to inhibit the growth of 0. This test is similar to that described in GB 2065653B; and (c) the test compound is expressed in cell culture in a human breast cancer cell line MC
Assay that measures the ability of F-7 to inhibit proliferation. This test was performed by Lippman et al. (Cancer Res., 1976,
36 , 4595).

The pharmacological properties of the quinazolines of the present invention will vary with changes in the structure, quinazolines of the present invention generally shows the thymidylate synthase inhibitory concentrations below: -IC _50,
For example 0.001-10 or 20 μM; or the quinazolines of the present invention show inhibition of the L1210 cell line at the following concentrations:
C ₅₀ , for example 0.001-50 or 100 μM.

Especially preferred quinazolines of the present invention, generally exhibit concentration thymidylate synthase inhibitory follows: -IC ₅₀ 1
or less; μM; or they show inhibition of the L1210 cell line at the following concentrations: −IC ₅₀ ≦ 10 μM.

For inhibition of the MCF-7 cancer cell line, the quinazolines of the present invention generally exhibit inhibitory activity at the following concentrations: -IC ₅₀₀ .
1-50 or -100 μM. Especially preferred quinazolines shows the MCF-7 cell line inhibitory concentrations below: -IC ₅₀ 5 [mu] M
Less than.

For example, quinazoline Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N -Methyl-L-glutamic acid has an IC _{50 of} 0.0095 μM against thymidylate synthase,
L1210 0.24 IC ₅₀ of relative cell lines, and shows an IC ₅₀ of 0.3μM against MCF-7 cell line.

In vivo studies in mice as assessed by measuring the amount of cleavage product as a percentage of the total amount of parent compound and cleavage product present in the liver or plasma of mice sacrificed 1 hour after intraperitoneal administration of the dipeptide 100 mg / kg for various compounds of the invention
At a dose of 4. showed substantially no cleavage of the central amide bond of the dipeptide. That is, the cleavage was within about 5%, which is the limit of detection by experiment. When it is used in practice, slight cleavage of this amide bond of the quinazolines is acceptable, but a level of cleavage of 10% or less in vivo upon administration is preferred, not more than about 5%, especially 2% or 1%. % Is desirable.

The quinazolines of the present invention can be administered to warm-blooded animals, including humans, in the form of a pharmaceutical composition comprising the quinazolines and a pharmaceutically acceptable diluent or carrier.

The compositions of the present invention may be in a form suitable for oral administration, such as tablets or capsules, or especially for parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular, or infusion), such as a sterile solution Suspensions or emulsions, or in a form suitable for topical administration, such as an ointment or cream, or in a form suitable for anal administration, such as a suppository.

These compositions may contain, in addition to the quinazolines of the present invention, for example, one or more anticancer agents selected from: mitotic inhibitors such as vinblastine; alkylating agents such as cisplatin, carboplatin, and cyclophosphamido. Other antimetabolites such as 5-fluorouracil, cytosine arabinoside, and oxyurea; antibiotics that cause intercalation;
For example, adriamycin and bleomycin; enzymes such as asparaginase; topoisomerase inhibitors such as etoposide; and biological response modifiers such as interferon.

In general, the composition of the present invention can be prepared by a conventional method using a common excipient.

Quinazolines of the present invention is usually m ² per 50-25000mg body area of the animal, i.e. at a dose ranging from about 1-500 mg / kg, is administered to a warm-blooded animal. However, doses outside this range can be used if desired, especially if the preferred mode of administration by subcutaneous injection is to be employed.
mg / kg can be increased. Preferably 10-150 mg / k
g, especially a daily dose of 30-80 mg / kg, is used.

Accordingly, the present invention provides a method of treating a patient, particularly a warm-blooded animal, eg, a human, in need of such treatment which assists in regression and alleviation of the cancer, comprising administering to said patient an effective amount of a quinazoline as defined above. And methods comprising: The invention also provides for the use of these quinazolines in the manufacture of a novel drug for use in treating cancer.

The quinazolines of the present invention are important in a wide range of antitumor activities, particularly in humans, including breast, uterine and liver cancers. Furthermore, they are important for the treatment of a certain range of leukemias, lymphocytic malignancies and solid tumors such as cancer and sarcoma.

For example, in view of the activities exhibited by the aforementioned antimetabolites, such as aminopterin and methotrexate,
The quinazolines of the present invention are also important for use in treating other conditions, such as allergic conditions, such as psoriasis. In using the quinazolines of the present invention for these purposes, these compounds are usually administered at doses in the range of 50-25000 mg / m ² of animal body area, ie, about 1-500 mg / kg. However, doses outside the range can be used if desired. Generally allergic conditions,
For example, for the treatment of psoriasis, topical administration of the quinazolines of the invention is preferred. For example, for topical administration a daily dose of, for example, 10-150 mg / kg, in particular 30-80 mg / kg, may be used.

The compositions containing the quinazolines of the present invention can be administered in a single dosage form, ie, in separate dosage forms, each containing a single dosage, or in multiples or submultiples of the single dosage, eg, 1-25.
It can be prepared with quinazoline amounts ranging from 0 or 500 mg.

 The invention is illustrated by the following example.

The structures of all compounds of the invention were confirmed by proton nuclear magnetic resonance and mass spectroscopy, and by elemental analysis. Proton nuclear magnetic resonance and mass spectroscopy were performed by Bruker (Br) operating at a field strength of 250 MHz.
uker) Measured using a WM250 spectrometer. Chemical shifts are ppm down from tetramethylsilane as back reference
(Δ scale), and thus the peak multiplicity is given by: s, singlet; d, doublet; d of d's, doublet doublet; t, triplet; m, multiplet Item: Assignments for each of the signals shown in the same manner are considered appropriate. Mass spectrum is ZAB for VG analysis by fast atom bombardment ionization (FAB)
Finnigan by SE spectrometer or electrospray ionization (ESI) or chemical ionization (CI)
igan) Obtained using a TSQ 700 spectrometer. Positive ion data or negative ion data were collected as appropriate.

Column chromatography was performed using Merck Art 15111 silica gel. Petroleum ether has a boiling point of 60-
It means a fraction at 80 ° C.

Other groups used in the production of the compound of the present invention, R ¹ , R ² ,
Intermediates having R ⁴ , R ⁸ and Ar are described in GB 2 065
653, 2 175 903, 2 188 319 and 2 202 84
No. 7 and UK Patent Application No. 2 217 709, 2 227
Nos. 016 and 2 244 708 and their corresponding applications filed in other countries.

Examples Example 1: Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid (1) tri-tert-butyl-L-γ-glutamyl -N-
Methyl-L-glutamate N-methyl-L-glutamic acid (3.2 g) in water (50 ml)
And the pH of the solution is adjusted to 9 using solid sodium bicarbonate.
Was adjusted. The solution was cooled to 0 ° C. and benzyl chloroformate (9 ml) was added dropwise with vigorous stirring over 20 minutes. The mixture was stirred at 0 ° C. for a further 2 hours and at room temperature for a further 2 hours, then diethyl ether (2 × 100 ml) was added.
Times). The product was then isolated by acidifying the aqueous solution to pH 3 with 5N HCl and extracting with ethyl acetate (2 × 100 ml). The pooled organic extracts were dried over anhydrous sodium sulfate and the solvent was concentrated in vacuo to give a yellow oil. It crystallized on standing. By recrystallization from water, N- (benzyloxycarbonyl) -N
-Methyl-L-glutamic acid (2.26 g) was obtained. Melting point 97
-98 ° C.

NMR spectrum: (CD ₃ SOCD ₃ , 67 ° C.) 1.94, 2.14 (2 × m, 2
H, β-CH ₂ ), 2.22 (t, 2H, J = 6.27Hz, γ-CH ₂ ), 2.79
(S, 3H, N-CH 3), 4.53 (dd, J = 5.0,10.4Hz, 1H, α-C
H), 5.08 (s, 2H , ArCH 2), 7.33 (m, 5H, ArH). Mass spectrum: (CI) m / e 296 (M + H) + Elemental analysis: Found C, 56.67; H, 5.78; N, 4.72% C 14 H 17 NO 6 theory C, 56.95; H, 5.80; N, 4.74 % N- (benzyloxycarbonyl) -N-methyl-L
Glutamic acid (1.85 g) was suspended in dichloromethane (52 ml) in a pressure bottle, to which concentrated sulfuric acid (0.28 ml) was added. The mixture was cooled to −20 ° C. and liquid isobutylene (25 ml) was added. Keep the stoppered reactor at room temperature for 28 hours,
Shake vigorously, then cool and add saturated sodium bicarbonate solution (100 ml) and ethyl acetate (200 ml). Separate the organic phase and add saturated sodium bicarbonate (2 x 160 ml)
And washed with water (100 ml), then dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography on a silica gel column using 10% ethyl acetate in dichloromethane as eluent. Thus, N- (benzyloxycarbonyl) -N-
Di-tert-butyl methyl-L-glutamate (1.8 g)
Was obtained as an oil.

NMR spectrum: (CD ₃ SOCD ₃ , 67 ° C.) 1.37, 1.38 (2 × s, 1
8H, C (CH ₃ ) ₃ ), 1.90,2.07 (2 xm, 2H, β-CH ₂ ), 2.20
(M, 2H, γ-CH ₂ ), 2.79 (s, 3H, N-CH ₃ ), 4.44 (dd, J =
5.3, 10.2Hz, 1H, α-CH), 5.11 (m, 2H, ArCH ₂ ), 7.33 (m,
5H, ArH). Mass spectrum: (CI) m / e 408 (M + H) + Elemental analysis: Found C, 64.74;, H, 8.15 ; N, 3.40% C 22 H 33 NO 6 theory C, 64.84; H, 8.16; N, 3.44% N- in ethyl acetate containing 10% Pd / C (0.22 g)
A solution of di-tert-butyl (benzyloxycarbonyl) -N-methyl-L-glutamate (1.6 g) was stirred under hydrogen for 2.5 hours. The catalyst was identified and the filtrate was concentrated to dryness in vacuo to give N-methyl-L-glutamic acid di-tert.
-Butyl (1.14 g) was obtained as an oil.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.39,1.42 (2 xs, 18H, C
(CH ₃ ) ₃ ), 1.67 (m, 2H, β-CH ₂ ), 2.19 (s, 3H, N-C
H ₃ ), 2.23 (m, 2H, γ-CH ₂ ), 2.91 (dd, J = 6.1, 7.8 Hz, 1
H, α-CH). Mass spectrum: (CI) m / e 274 (M + H) ⁺ α-tert-butyl-N-benzyloxycarbonyl-
L-glutamate (Org. Prep. Proc. Int., 1985, 17 , 416;
To a solution of (1.31 g) and N-methylmorpholine (0.37 g) in dry tetrahydrofuran (5 ml) (cooled to −20 ° C.) was added, with stirring, isobutyl chloroformate (0.50 g). After 10 minutes, di-tert-butyl N-methyl-L-glutamate (1.003 g) in tetrahydrofuran (5 m
The solution in l) was added. Stirring was continued at −20 ° C. for 10 minutes, then at laboratory temperature for 4 hours. The N-methylmorpholine hydrochloride was filtered off and the filtrate was concentrated in vacuo. The residue was purified by chromatography on a silica gel column using a gradient of ethyl acetate in dichloromethane (5-30% v / v) as eluent. The product was triturated in diethyl ether and the white solid was isolated by filtration,
Washed with gasoline and dried in vacuum. Thus, tri-tert-butyl N-methyl- [N- (benzyloxycarbonyl) -L-γ-glutamyl] -L-glutamate (1.62 g) was obtained. 92-93 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.32,1.37,1.38 (3 × s, 2
7H, C (CH ₃ ) ₃ ), 1.86,2.03 (2 xm, 4H, β-CH ₂ ), 2.1
2,2.38 (2 xm, 4H, γ -CH 2), 2.60,2.79 (2 xs, 3H, N-
CH ₃ ), 3.94 (m, 1H, glu α-CH), 4.49, 4.76 (2 x dd, J
= 4.8,10.8Hz, 1H, N-Me glu α-CH), 5.03 (m, 2H, ArCH
₂ ), 7.35 (s, 5H, ArH), 7.64 (t, J = 7.7Hz, 1H, NH). Mass spectrum: (CI) m / e 593 (M + H) + Elemental analysis: Found C, 62.84; H, 8.16; N, 4.69% C 31 H 48 N 2 O 9 theory C, 62.82; H, 8.16; N , 4.73% Tri-t in ethyl acetate containing 10% Pd / C (0.11 g)
A solution of ert-butyl N-methyl- [N (benzyloxycarbonyl) -L-γ-glutamyl] -L-glutamate (0.80 g) was stirred under hydrogen for 3 hours. The catalyst was filtered off and the filtrate was concentrated in vacuo to give tri-tert-butyl-L-γ-glutamyl-N-methyl-L-glutamate (0.61 g) as an oil.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.38, 1.39, 1.41, 1.42 (4
_{xs, 27H, C (CH 3} ) 3, 1.79,2.05 (2 xm, 4H, β-CH 2), 2.
12,2.39 (2 xm, 4H, γ -CH 2), 2.60,2.84 (2 xs, 3H, N
−CH ₃ ), 3.18 (dd, J = 8.3, 4.9 Hz, 1H, CHNH ₂ ), 4.53, 4.78
(2 x dd, J = 10.5, 4.6 Hz, 1H, N-Me glu α-CH). Mass spectrum: (CI) m / e 459 (M + H) ⁺ (2) Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoic acid tert-butyl p-aminobenzoate (Synth. Commun.,
1984, 14 , 921; 10.5 g), a mixture of propargyl bromide (7.3 ml of an 80% solution in toluene), potassium carbonate (7.5 g) and N, N-dimethylacetamide (85 ml) at 50 ° C.
For 24 hours, cooled, filtered and evaporated. The residue was purified on a silica gel column with 6: 1 hexane and ethyl acetate.
The v / v mixture was purified by chromatography using as eluent.

The above product (7.3 g), 6-bromomethyl-3,4-dihydro-2-methylkiazolin-4-one (8 g; prepared as described in Example 3 of GB 2 188 319B), calcium carbonate ( 3.2 g) and dimethylformamide (100 ml) was stirred at laboratory temperature for 65 hours, filtered and evaporated. The residue was purified by chromatography on a silica gel column using ethyl acetate as eluent.

The above product (2.5 g) and trifluoroacetic acid (25 m
The mixture of l) was stirred at laboratory temperature for 10 minutes and evaporated to give the above p-aminobenzoic acid as its trifluoroacetate salt (2.5 g).

(3) Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid Np- [N- (3,4-dihydro- 2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2
-Inyl) amino] benzoic acid trifluoroacetate (0.46
1g) and tri-tert-butyl-L-γ-glutamyl-N-methyl-L-glutamate (0.60 g) were dissolved in dry dimethylformamide (15 ml) at 0 ° C., and diethyl cyanophosphonate was added to the solution. (0.359 g) and then triethylamine (0.222 g) were added under a nitrogen atmosphere. The solution was stirred at 0 ° C. in the dark for 20 minutes, then at laboratory temperature for 2 hours, then diluted with ethyl acetate (100 ml) and water (100 ml). The aqueous phase was separated and extracted with ethyl acetate (2x100ml). The combined ethyl acetate extracts were combined with 10% aqueous citric acid (2 × 50 ml), saturated sodium bicarbonate (100 ml) and dilute aqueous sodium chloride (1 ml).
00 ml), then dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by chromatography on a silica gel column using ethyl acetate as eluent. The product is precipitated from dichloromethane / petroleum ether and tri-tert-butyl-Np-
[N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methyl-L-glutamate (0.56 g). Melting point 110-11
3 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.32,1.33,1.35,1.36,1.3
9 (5 xs, 27H, C (CH 3) 3), 1.86 (m, 4H, β-CH 2), 2.3
2 (s, 3H, quinazoline 2-CH ₃ ), 2.15,2.44 (2 xm, 4H,
γ-CH ₂ ), 2.61, 2.80 (2 xs, 3H, N-CH ₃ ), 3.23 (s, 1H,
C≡CH), 4.29 (m, 3H, CH ₂ C≡C, glu α-CH), 4.48, 4.77
(2 xm, 3H, quinazoline _{6-CH 2, N-Me} glu α-CH),
6.82 (d, J = 8.7 Hz, 2H, 3 ', 5'-ArH), 7.54 (d, J = 8.4H
z, 1H, quinazoline 8-H), 7.71 (t, J = 8.6 Hz, 3H, quinazoline 7-H, 2 ', 6'-ArH), 7.96 (s, 1H, quinazoline 5-H), 8.28 (t , 1H, glu NH), 12.19 (s, 1H lactam
NH). Mass spectrum: (cationic FAB) m / e 810 (M + Na)
⁺ Elemental Analysis: Found C, 64.75; H, 7.19; N, 8.73% C 43 H 57 N 5 O 0 · 0.5H 2 O Theoretical value C, 64.81; H, 7.33; N, 8.79% tri -tert- butyl -Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N- A mixture of methyl-L-glutamate (0.167 g) and trifluoroacetic acid (10 ml) was stirred at laboratory temperature in the dark for 1 hour. The solution was then concentrated in vacuo and the residue was triturated with diethyl ether (30ml). The white solid was isolated by filtration, washed with diethyl ether (4 × 10 ml) and dried in vacuo. Thus Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N- Methyl-L-glutamic acid trifluoroacetate (0.152 g) was obtained. Melting point 205 ° C, decomposition.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.80-2.14 (m, 4H, β-CH
_{2), 2.42 (s, 3H} , quinazoline _{2-CH 3), 2.14,2.45 (} 2
xm, 4H, γ-CH ₂ ), 2.65,2.81 (2 xs, 3H, N-CH ₃ ), 3.24
(S, 1H, C≡CH), 4.35 (s, 3H, CH 2 C≡C, glu α-CH), 4.8
1 (s, 2H, quinazoline _{6-CH 2), 4.58,4.91 (} dd, J = 11.
0,4.5Hz, 1H, N-Me glu α-CH), 6.83 (d, J = 8.7Hz, 2H,
3 ', 5'-ArH), 7.59 (d, J = 8.4Hz, 1H, quinazoline 8
-H), 7.75 (m, 3H, 2 ', 6'-ArH, quinazoline 7-
H), 8.00 (s, 1H, quinazoline 5-H), 8.32 (d, J = 7.
3Hz, 1H, glu NH). Mass spectrum: (cationic FAB) m / e 642 (M + Na)
⁺ Elemental Analysis: Found C, 52.93; H, 5.18; N, 8.41; F, 7.65% C 31 H 33 N 5 O 9 · 1.1CF 3 COOH · 0.5Et 2 O · 1H 2 O Theoretical value C, 52.84; H, 5.18; N, 8.75; F, 7.83% Example 2: Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methylglycine (1) di-tert-butyl-L-γ-glutamyl-N- Methylglycine N-methylglycine (7.0 g) was added to water (50 ml),
The pH of the solution was adjusted to 9 with solid sodium bicarbonate. The solution was cooled to 0 ° C. and benzyl chloroformate (19 ml) was added dropwise with vigorous stirring over 20 minutes. The mixture was stirred at 0 ° C. for a further 2 hours and at room temperature for a further 2 hours, then extracted with diethyl ether (2 × 130 ml). The aqueous solution was then acidified to pH 3 with 5N HCl and the product was isolated by extraction with ethyl acetate (2 × 130 ml). The combined organic extracts were dried over anhydrous sodium sulfate and the solution was concentrated in vacuo to give N- (benzyloxycarbonyl) -N-methylglycine as a pale yellow oil (16.2g).

NMR _{Spectrum: (CD 3 SOCD 3, 72} ℃) 2.91 (s, 3H, N-C
_{H 3), 3.94 (s,} 2H, CH 2 CO 2 H), 5.07 (s, 2H, ArCH 2), 7.33
(S, 5H, ArH). Mass spectrum: (ESI) m / e 233 (M ⁺ ) N- (benzyloxycarbonyl) -N-methylglycine (7.86 g) and tert-butyl alcohol (4.68 m)
To a solution of l) in chloroform (30 ml) (cooled in an ice-water bath), under stirring, add a solution of dicyclohexylcarbodiimide (8.0 g) in chloroform (30 ml), then add 4-dimethylamino as catalyst Pyridine (0.04 g) was added. Stirring was continued at 0 ° C. for 2 hours, then the mixture was kept at 4 ° C. overnight. The dicyclohexylurea was filtered off and the solvent was concentrated in vacuo. The residue was dissolved in ethyl acetate (110ml) and acetic acid (1.10ml) was added. The solution was kept at 4 ° C. for 2 hours. After filtration, ethyl acetate was added with 5% aqueous sodium bicarbonate solution (2 times with 70 ml) and water (3 times with 80 ml).
) And dried over anhydrous magnesium sulfate. The solution was concentrated in vacuo and the residue was purified by chromatography on a silica gel column using 30% ethyl acetate in gasoline as eluent. Thus tert-butyl-N
-(Benzyloxycarbonyl) -N-methylglycine (3.6 g) was obtained as an oil.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.36,1.41 (2 xs, 9H, C
(CH ₃ ) ₃ ), 2.88, 2.91 (2 xs, 3H, N-CH ₃ ), 3.91, 3.94
_{(2 xs, 2H, CH 2} COOH), 5.05,5.10 (2 xs, 2H, ArCH 2),
7.31, 7.36 (2 xm, 5H, ArH). Mass spectrum: (ESI) m / e 279 (M ⁺ ) Elemental analysis: found C, 64.27; H, 7.62; N, 5.03% C ₁₅ H ₂₁ NO ₄ theoretical C, 64.50; H, 7.58; N, 5.01 % Tert- in ethyl acetate containing 10% Pd / C (0.23 g)
A solution of butyl-N- (benzyloxycarbonyl) -N-methylglycine (1.60 g) was stirred under hydrogen for 2.5 hours. The catalyst was filtered off and the filtrate was concentrated in vacuo to give tert-butyl-N-methylglycine (0.71 g) as an oil.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.41 (s, 9H, C (C
_{H 3) 3), 2.25 (} s, 3H, N-CH 3), 3.13 (s, 2H, CH 2). Mass spectrum: (CI) m / e 146 (M + H) ⁺ α-tert-butyl-N-benzyloxycarbonyl-
To a solution of L-glutamate (1.63 g) and N-methylmorpholine (0.487 g) in dry tetrahydrofuran (8 ml) (cooled to -20 ° C) was added, with stirring, isobutyl chloroformate (0.657 g). After 10 minutes, a solution of tert-butyl-N-methylglycine (0.70 g) in tetrahydrofuran (8 ml) was added. Stirring was continued at −20 ° C. for 10 minutes, then at laboratory temperature for 4 hours. The N-methylmorpholine hydrochloride was filtered off and the filtrate was concentrated in vacuo. The residue is purified by chromatography on a silica gel column using 5% ethyl acetate in dichloromethane as eluent, di-tert-butyl-N-methyl- [N- (benzyloxycarbonyl) -L-γ-glutamine] glycine Was obtained as a yellow oil (1.73 g).

NMR spectrum: (CD ₃ SOCD ₃ ) 1.32,1.38,1.39,1.41, (4
xs, 18H, C (CH ₃ ) ₃ ), 1.77,1.87 (2 xm, 2H, β-C
H ₂ ), 2.24, 2.38 (2 xm, 2H, γ-CH ₂ ), 2.79, 2.95 (2 x
_{s, 3H, N-CH 3} ), 3.93 (m) and 4.08 (s) (3H, CH 2 CO
^{_{2 Bu t, α-CH)}} , 5.03 (m, 2H, ArCH 2), 7.36 (m, 5H, ArH),
7.64 (t, 1H, NH). Mass spectrum: (CI) m / e 465 (M + H) + Elemental analysis: Found C, 61.56; H, 7.81; N, 5.97% C 24 H 36 N 2 O 7 · 0.5H 2 O Theoretical value C, 61.45; H, 7.84; N, 5.97% Di-te in ethyl acetate containing 10% Pd / C (0.150 g)
A solution of rt-butyl-N-methyl- [N- (benzyloxycarbonyl) -L-γ-glutamine] glycine (0.80 g) was stirred under hydrogen for 6 hours. The catalyst was filtered off and the filtrate was concentrated in vacuo to give di-tert-butyl-L-γ-glutamyl-N-methylglycine (0.53 g) as a yellow oil.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.40, 1.41, 1.43 (3 xs, 1
_{8H, C (CH 3) 3} ), 1.59,1.77 (2 xm, 2H, β-CH 2), 2.2
4,2.40 (2 xm, 2H, γ -CH 2), 2.79,2.99 (2 xs, 3H, N-
CH ₃ ), 3.17 (m, 1H, α-CH), 4.02 (s) and 4.10
_{(D) (2H, CH 2} CO 2 Bu t). Mass spectrum: (ESI) m / e 331 (M + H) ⁺ (2) Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methylglycine Np- [N- (3,4-dihydro-2-methyl) -4-
Oxoquinazolin-6-ylmethyl) -N- (prop-
2-ynyl) amino] benzoic acid trifluoroacetate (0.
553 g) and di-tert-butyl-L-γ-glutamyl-
A mixture of N-methylglycine (0.53 g) was dissolved in dry dimethylformamide (15 ml) at 0 ° C. and to this solution was added diethyl cyanophosphonate (0.359 g) followed by triethylamine (0.222 g) under a nitrogen atmosphere. . The solution was stirred at 0 ° C. in the dark for 20 minutes, then at laboratory temperature for 3 hours, then ethyl acetate (100 ml) and water (100 ml)
l). The aqueous phase was separated and ethyl acetate (2 in 100 ml)
Times) extracted. The combined ethyl acetate extracts were combined with a 10% aqueous citric acid solution (50 ml twice), saturated sodium bicarbonate (100 m
l) and diluted aqueous sodium chloride solution (100 ml),
It was then dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by chromatography on a silica gel column using ethyl acetate as eluent. The product is precipitated from cold dichloromethane / diethyl ether, thus di-tert-butyl-Np- [N-
(3,4-Dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methylglycine (0.22 g) was obtained. Was. 104-108 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.30, 1.39 (2 xs, 18H, C
_{(CH 3) 3), 2.33} (s, 3H, quinazoline 2-CH _3), 1.96
(M, 2H, β-CH ₂ ), 2.26,2.45 (2 xm, 2H, γ-CH ₂ ), 2.7
9,2.95 (2 xs, 3H, N -CH 3), 3.23 (s, 1H, C≡CH), 3.95
(D) and 4.06 (s) (2H, CH ₂ CO ₂ ), 4.25 (m, 1H, α
-CH), 4,33 (s, 2H , CH 2 C≡C), 4.77 (s, 2H, quinazoline _{6-CH 2), 6.81 (} m, 2H, 3 ', 5'-ArH), 7.54 (d , J =
8.3Hz, 1H, quinazoline 8-H), 7.74 (m, 3H, quinazoline 7-H, 2 ', 6'-ArH), 7.96 (s, 1H, quinazoline 5-H)
H), 8.28 (t, 1H, NH), 12.19 (s, 1H, lactam NH). Mass spectrum: (cationic FAB) m / e 682 (M + Na)
⁺ Elemental Analysis: Found C, 65.12; H, 6.86; N, 10.50% C 36 H 45 N 5 O 7 · 0.25H 2 O Theoretical value C, 65.09; H, 6.90; N, 10.54% di -tert- butyl -Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N- Methyl glycine (0.074g)
And a mixture of trifluoroacetic acid (5 ml) was stirred at laboratory temperature in the dark for 1 hour. The solution was then concentrated in vacuo and the residue was triturated with diethyl ether (20ml). The white solid was isolated by filtration, washed with diethyl ether (4 × 10 ml) and dried in vacuo. Thus Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methylglycine trifluoroacetate (0.60
g) was obtained. 140 ° C, decomposes.

NMR spectrum (CD ₃ SOCD ₃ ) 2.01 (m, 2H, β-CH ₂ ), 2.3
3,2.42 (2 xm, 2H, γ-CH ₂ ), 2.42 (s, 3H, quinazoline
_{2-CH 3), 2.81,2.97 (} 2 xs, 3H, N-CH 3), 3.24 (s, 1H,
C≡CH), 3.98 (d, J = 8.0 Hz) and 4.10 (s) (2H, CH ₂
CO ₂ ), 4.36 (m, 3H, CH ₂ C≡C, α-CH), 4.81 (s, 2H, quinazoline 6-CH ₂ ), 6.84 (d, J = 8.3 Hz, 2H, 3 ', 5' −Ar
H), 7.59 (d, J = 8.5 Hz, 1H, quinazoline 8-H), 7.76
(M, 3H, quinazoline 7-H, 2 ', 6'-ArH), 8.01 (s, 1
H, quinazoline 5-H), 8.33 (t, 1H, NH). Mass spectrum: (cationic FAB) m / e 570 (M + Na)
⁺ Elemental Analysis: Found C, 53.98; H, 5.63; N, 9.02; F.8.15% C 28 H 29 N 5 O 7 · 1.1CF 3 COOH · 0.8H 2 O · 0.8Et 2 O Theoretical value C, 53.72 H, 5.35; N, 9.38; F, 8.39% Example 3: Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-L-proline (1) di-tert-butyl-L-γ-glutamyl-L-proline α-tert-butyl-N-benzyloxycarbonyl-
To a solution of L-glutamate (3.15 g) and N-methylmorpholine (0.909 g) in dry tetrahydrofuran (10 ml) (cooled to -20 ° C) was added, with stirring, isobutyl chloroformate (1.22 g). After 10 minutes, a solution of tert-butyl-L-proline (1.54 g) in tetrahydrofuran (10 ml) was added. Stirring was continued at −20 ° C. for 10 minutes, then at laboratory temperature for 4 hours. The N-methylmorpholine hydrochloride was filtered off and the filtrate was concentrated in vacuo. Residue on silica gel column 10% to 30% in dichloromethane
Purification by chromatography using a gradient of ethyl acetate as eluent. The product was triturated in gasoline and a white solid was isolated by filtration, washed with gasoline and dried in vacuo. Thus di-tert-butyl-
N- [N-benzyloxycarbonyl) -L-γ-glutamyl] -L-proline (2.86 g) was obtained. Melting point 106-10
8 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.37, 1.39 (2 xs, 18H, C
(CH ₃ ) ₃ ), 1.62-2.10 (m, 6H, glu β-CH ₂ , proline 3- and 4-CH ₂ ), 2.33 (m, 2H, glu γ-CH ₂ ), 3.42
(M, 2H, proline _{5-CH 2), 3.93 (} m, 1H, glu α-CH),
4.13, 4.40 (2 x dd, J = 8.8,3.9Hz, 1H, proline 2-C
H), 5.03 (m, 2H, ArCH ₂ ), 7.35 (m, 5H, ArH), 7.59 (t, 1
H, glu NH). Mass spectrum: (CI) m / e 491 (M + H) + Elemental analysis: Found C, 63.39; H, 7.78; N, 5.69% C 26 H 38 N 2 O 7 theory C, 63.65; H, 7.81; N , 5.71% 10% Pd / C (0.15 g) in ethyl acetate containing 0.15 g
t-butyl-N- [N- (benzyloxycarbonyl)
-L-γ-glutamyl] -L-proline (0.98 g) was stirred under hydrogen for 8 hours. The catalyst was filtered off and the filtrate was concentrated in vacuo to give di-tert-butyl-L-γ-glutamyl-L-proline (0.70 g) as an oil.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.37, 1.40 (2 xs, 18H, C
(CH ₃ ) ₃ ), 1.55-2.10 (m, 6H, glu β-CH ₂ , proline 3- and 4-CH ₂ ), 2.33 (t, J = 7.5 Hz, 2H, γ-CH ₂ ),
3.16 (m, 1H, glu α-CH), 3.48 (t, J = 6.7 Hz, 2H, proline 5-CH ₂ ), 4.14, 4.40 (2 × dd, J = 8.8,3.9 Hz, 1H, proline 2- CH). Mass spectrum: (CI) m / e 357 (M + H) ⁺ (2) Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-L-proline Np- [N- (3,4-dihydro-2-methyl- 4-
Oxoquinazolin-6-ylmethyl) -N- (prop-
2-ynyl) amino] benzoic acid trifluoroacetate (0.
461 g) and di-tert-butyl-L-γ-glutamyl-
A mixture of L-proline (0.524 g) was dissolved in dry dimethylformamide (15 ml) at 0 ° C. and to this solution was added diethyl cyanophosphonate (0.359 g) followed by triethylamine (0.222 g) under a nitrogen atmosphere. Bring this solution to 0 ° C
For 20 minutes in the dark and then for 3 hours at laboratory temperature, then diluted with ethyl acetate (100 ml) and water (100 ml). The aqueous layer was separated and extracted with ethyl acetate (2x100ml). The combined ethyl acetate extracts were combined with a 10% aqueous citric acid solution (50 ml twice), saturated sodium bicarbonate (100 ml)
And washed with dilute aqueous sodium chloride solution (100 ml), then dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography on a silica gel column using ethyl acetate as eluent.
The product is precipitated from cold diethyl ether / gasoline,
Thus di-tert-butyl-N-p- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L- γ-glutamyl-L-proline (0.192 g) was obtained. 113-117 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.31,1.35,1.39 (3 × s, 1
8H, C (CH ₃ ) ₃ ), 1.70-2.10 (m, 6H, glu β-CH ₂ , proline 3- and 4-CH ₂ ), 2.32 (s, 3H, quinazoline 2-
CH ₃ ), 2.38 (t, J = 7.5 Hz, 2H, glu γ-CH ₂ ), 3.19 (s, 1
H, C≡CH), 3.41 (m, 2H, proline 5-CH ₂ ), 4.15, 4.40
(2 x dd, J = 9.0, 3.8 Hz, 1H, proline 2-CH), 4.32 (m,
3H, CH ₂ C≡C, glu α-CH), 4.77 (s, 2H, quinazoline 6
−CH ₂ ), 6.83 (d, J = 8.9Hz, 2H, 3 ′, 5′-ArH), 7.53
(D, J = 8.4 Hz, 1H, quinazoline 8-H), 7.68 (dd, J =
8.4Hz, 2.0Hz, 1H, quinazoline 7-H), 7.73 (d, J = 9.0
Hz, 2H, 2 ', 6'-ArH), 7.97 (d, J = 2.0 Hz, 1H, quinazoline 5-H), 8.23 (t, 1H, NH). Mass spectrum: (cationic FAB) m / e 708 (M + Na)
⁺ Elemental Analysis: Found C, 65.81; H, 6.88; N, 10.00% C 38 H 47 N 5 O 7 · 0.5H 2 O Theoretical value C, 65.69; H, 6.96; N, 10.08% di -tert- butyl -Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-L- A mixture of proline (0.067 g) and trifluoroacetic acid (5 ml) was stirred at laboratory temperature in the dark for 1 hour. The solution was then concentrated in vacuo and the residue was triturated with diethyl ether (20ml). The white solid was isolated by filtration, washed with diethyl ether (4 × 10 ml) and dried in vacuo. Thus Np- [N- (3,4-dihydro-2-methyl-4-
Oxoquinazolin-6-ylmethyl) -N- (prop-
2-Inyl) amino] benzoyl-L-γ-glutamyl-L-proline trifluoroacetate (0.054 g) was obtained. 162 ° C, decomposes.

NMR spectrum: (CD ₃ SOCD ₃ 1.84,2.08 (2 xm, 6H, glu
β-CH ₂ , proline 3- and 4-CH ₂ ), 2.39 (m, 5H, quinazoline 2-CH ₃ , glu γ-CH ₂ ), 3.23 (s, 1H, C≡C
H), 3.42 (m, 2H , proline _{5-CH 2), 4.34 (} m, 3H, CH 2 ≡
C, glu α-CH), 4.23, 4.48 (dd, J = 9.9, 3.5 Hz, 1H, proline 2-CH), 4.80 (s, 2H, quinazoline 6-CH ₂ ), 6.83
(D, J = 8.8 Hz, 2H, 3 ', 5'-ArH), 7.57 (d, J = 8.4 Hz, 1
H, quinazoline 8-H), 7.74 (d, J = 8.7 Hz, 3H, 2 ', 6'
-ArH, quinazoline 7-H), 7.99 (s.1H, quinazoline
5-H), 8.32 (t, 1H, NH). Mass spectrum: (cationic FAB) m / e 596 (M + Na)
⁺ Elemental Analysis: Found C, 55.44; H, 4.91; N, 9.81% C 30 H 31 N 5 O 7 · 1.1CF 3 COOH · 0.22Et 2 O Theoretical value C, 55.54; H, 4.83; N, 9.79% Example 4: Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-α-methylalanine (1) di-tert-butyl-L-γ-glutamyl-α- Methylalanine α-aminobutyric acid (6.18 g), tert-butyl acetate (200 m
l) and a 70% aqueous solution of perchloric acid (9.45 g) were stirred at laboratory temperature for 7 days. The mixture was then cooled in an ice-water bath and
Extracted with 5N hydrochloric acid (4 times with 50 ml). The aqueous extracts were combined and immediately neutralized with solid sodium bicarbonate. The aqueous solution was extracted with diethyl ether (3 × 100 ml), the ether extracts were pooled, dried over anhydrous sodium sulfate, and the ether was evaporated in vacuo to give tert-butyl-α-methylalanine (4.98 g). I got

NMR spectrum: (CD ₃ SOCD ₃ ) 1.16 (s, 6H, C (C
_{H 3) 2), 1.40 (} s, 9H, C (CH 3) 3). α-tert-butyl-N-benzyloxycarbonyl-
To a solution of L-glutamate (3.033 g) and N-methylmorpholine (0.909 g) in dry tetrahydrofuran (10 ml) (cooled to -20 ° C) was added, with stirring, isobutyl chloroformate (1.224 g). After 10 minutes, a solution of tert-butyl-α-methylalanine (2.0 g) in tetrahydrofuran (5 ml) was added. Stirring was continued at −20 ° C. for 10 minutes, then at laboratory temperature for 1 hour. The N-methylmorpholine hydrochloride was filtered off and the filtrate was concentrated in vacuo. The residue was purified by chromatography on a silica gel column using 2% methanol in dichloromethane as eluent. The product is crystallized from diethyl ether / gasoline and the white solid is isolated by filtration, washed with gasoline,
Dry in vacuum. Thus di-tert-butyl- [N
-(Benzyloxycarbonyl) -L-γ-glutamyl] -L-α-methylalanine (3.855 g) was obtained. Melting point
109-110 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.27 (s, 6H, C (C
_{H 3) 2), 1.34,1.39 (} 2 xs, 18H, C (CH 3) 3), 1.73,1.
90 (2 xm, 2H, β-CH ₂ ), 2.13 (t, J = 7.3 Hz, 2H, γ-C
H ₂ ), 3.90 (m, 1H, glu α-CH), 5.03 (m, 2H, ArCH ₂ ), 7.
36 (m, 5H, ArH), 7.64 (d, J = 7.5Hz, 1H, glu NH), 8.06
(S, 1H, α-methylalanine NH). Mass spectrum: (cationic FAB) m / e 501 (M + Na)
⁺ Elemental Analysis: Found C, 62.73; H, 8.02; N, 5.82% C 25 H 38 N 2 O 7 theory C, 62.74; H, 8.00; N, 5.85% 10% Pd / C a (0.2 g) A solution of the above product (0.669 g) in ethyl acetate was stirred under hydrogen for 3 hours. The catalyst was filtered off and the filtrate was concentrated in vacuo to give di-tert-butyl-L-γ-glutamyl-α-methylalanine (0.541 g).
Was obtained as an oil.

(2) Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-α-methylalanine Using the method described in Example 1 (3) as a starting material, tert-butyl-L-γ-glutamyl-N-methyl-
Di-tert-butyl-L-γ instead of L-glutamate
-Repeated with glutamyl-α-methylalanine (0.541 g). Thus di-tert-butyl-Np- [N-
(3,4-Dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-α-methylalanine (0.486 g) was obtained. Was. 115-116 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.26 (s, 6H, C (C
_{H 3) 2), 1.33,1.39 (} 2 xs, 18H, C (CH 3) 3), 1.91,1.
99 (2 xm, 2H, β-CH ₂ ), 2.14 (m, 2H, γ-CH ₂ ), 2.32
(S, 3H, quinazoline _{2-CH 3), 3.27 (} s, 1H, C≡CH), 4.
25 (m, 1H, glu α-CH), 4.34 (s, 2H, CH ₂ C≡C) 4.78
(S, 2H, quinazoline _{6-CH 2), 6.83 (} d, J = 8.9Hz, 2H,
3 ', 5'-ArH), 7.54 (d, J = 8.4Hz, 1H, quinazoline 8
-H), 7.70 (dd, J = 2.0Hz, 1H, quinazoline 7-H),
7.74 (d, J = 8.8 Hz, 2H, 2 ', 6'-ArH), 7.96 (d, J = 1.6H
z, 1H, quinazoline 5-H), 8.07 (s, 1H, α-methylalanine NH), 8.34 (d, J = 7.4 Hz, 1H, glu NH), 12.20 (s, 1
H, lactam NH). Mass spectrum: (cationic FAB) m / e 674 (M + H)
⁺ Elemental Analysis: Found C, 65.05; H, 7.04; N, 10.18% C 37 H 47 N 5 O 7 · 0.5H 2 O Theoretical value C, 65.08; H, 7.09; N, 10.26% di -tert- butyl -Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-glutamyl-α- Methylalanine (0.1 g) was treated with trifluoroacetic acid as described in Example 1 (3). Thus Np- [N- (3,4-dihydro-2-
Methyl-4-oxoquinazolin-6-ylmethyl) -N
-(Prop-2-ynyl) amino] benzoyl-L-γ
-Glutamyl-α-methylalanine trifluoroacetate (0.062 g) was obtained. 155-158 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.29 (s, 6H, C (C
_{H 3) 2), 1.89,2.02 (} 2 xm, 2H, β-CH 2), 2.18 (m, 2H,
γ-CH ₂ ), 2.38 (s, 3H, quinazoline 2-CH ₃ ), 3.23
(S, 1H, C≡CH), 4.29 (m, 1H, glu α-CH), 4.34 (s, 2H,
CH ₂ C≡C), 4.80 (s, 2H, quinazoline 6-CH ₂ ), 6.83
(D, J = 8.3 Hz, 2H, 3 ', 5'-ArH), 7.57 (d, J = 8.3 Hz, 1
H, quinazoline 8-H), 7.75 (d, J = 8.1 Hz, 3H, 2 ', 6'
-ArH and quinazoline 7-H), 7.98 (s, 1H, quinazoline 5-H), 8.02 (s, 1H, α-methylalanine NH),
8.29 (d, J = 7.6 Hz, 1 H, glu NH), 12.40 (br, CO ₂ H). Mass spectrum: (cationic FAB) m / e 584 (M + Na)
⁺ Elemental Analysis: Found C, 55.21; H, 5.00; N, 10.57; F, 5.80% C 20 H 31 N 5 O 7 · 0.7CF 3 COOH · 1H 2 O Theoretical value C, 55.37; H, 5.15; N , 10.62; F, 6.05% Example 5: Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-4-methylglutamyl-L-glutamic acid (1) α-tert-butyl-γ-methyl-N- Trityl-
L-4-methylglutamate γ-methyl-L-glutamate (12.88 g), tert-acetic acid
-Butyl (500 ml) and a 70% aqueous solution of perchloric acid (12.6
g) was stirred at room temperature for 88 hours. The mixture was then cooled in an ice-water bath and extracted with cold 0.5N hydrochloric acid (3 × 130 ml). The combined aqueous extracts were immediately neutralized with solid sodium bicarbonate and then extracted with diethyl ether (3 × 200 ml). The combined ether extracts were dried over anhydrous sodium sulfate, filtered, and the solution was concentrated in vacuo to
α-tert-butyl-γ-methyl-L-glutamate (1
4.5 g) were obtained as a colorless oil.

Α-tert-butyl-γ in dry chloroform (50 ml)
To a solution of -methyl-L-glutamate (13.71 g) was added, with stirring, triethylamine (8.49 g), followed by a solution of lead nitrate (8.34 g) and then a solution of trityl chloride (11.67 g) in dry chloroform (20 ml). Was added. 18 at room temperature
After stirring for an hour, a white solid precipitated from the solution. This was filtered off, the filtrate was concentrated in vacuo and the orange residue was purified by chromatography on a silica gel column using a gradient of dichloromethane in hexane (10-100%) as eluent. Thus, α-tert-butyl-γ-methyl-N-trityl-L-glutamate (9.46 g) was obtained as an oil. This solidified when allowed to stand at -20 ° C for 4 weeks. 79-80 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.13 (s, 9H, C (C
_{H 3) 3), 1.80,1.92 (} 2 xm, 2H, β-CH 2), 2.18,2.44
(2 xm, 2H, γ-CH ₂ ), 2.79 (d, J = 9.0 Hz, 1H, NH), 3.17
(M, 1H, α-CH), 3.60 (s, 3H, OCH ₃ ), 7.16-7.41 (m, 15
H, TrH). Mass spectrum: (CI) m / e 460 (M + H) + Elemental analysis: Found C, 76.13; H, 7.29; N, 2.89% C 29 H 33 NO 4 theory C, 75.79; H, 7.24; N, 3.05 0.6M potassium in tetrahydrofuran (13.75ml)
To a solution of hexamethyldisilazide at -78 ° C with stirring under argon was added α- in tetrahydrofuran (25 ml).
A solution of tert-butyl-γ-methyl-N-trityl-L-glutamate (2.52 g) was added over 5 minutes.
The resulting yellow solution was stirred at -78 C for 50 minutes, then methyl iodide (1.56 g) was added. Stirring was continued at −78 ° C. for 1.25 hours, then the white slurry was poured into a saturated ammonium chloride solution (200 ml). The mixture was extracted with diethyl ether (2 × 200 ml), combined with ether extractive drying over anhydrous sodium sulfate, filtered, and the solution was concentrated in vacuo. The residue was purified by chromatography on a silica gel column using a gradient of ethyl acetate in hexane (3-9%) as eluent. Thus, α-tert-butyl-γ-methyl-N-trityl-
L-4-methylglutamate (1.62 g) was obtained as a viscous oil. It was a mixture of two diastereoisomers (2S, 4S and 2S, 4R).

NMR spectrum: (CD ₃ SOCD ₃ ) 0.98, 1.04 (2 xd, J = 7.0
Hz, 3H, γ-CH ₃ ), 1.12, 1.14 (2 xs, 9H, C (CH ₃ ) ₃ ),
1.38,1.54 (2 xm, 1H, β-CH), 1.93,2.10 (2 xm, 1H,
β-CH), 2.40 (m, 1H, γ-CH), 2.75, 2.80 (2 xd, J ₁ =
_{8.9Hz, J 2 = 9.4Hz, 1H} , NH), 3.10 (m, 1H, α-CH), 3.50,
3.66 (2 xs, 3H, CO 2 CH 3), 7.16-7.39 (m, 15H, TrH). Mass spectrum: (CI) m / e 474 (M + H) + Elemental analysis: Found C, 76.12; H, 7.61; N, 2.89% C 30 H 35 NO 4 theory C, 76.08; H, 7.45; N, 2.96 % (2) Tri-tert-butyl-L-γ-4-methylglutamyl-L-glutamate To a mixture of lithium hydroxide monohydrate (0.117 g) in water (1.86 ml) was added tetrahydrofuran (27 ml). Of α-tert-butyl-γ-methyl-N-trityl-L-4-methylglutamate (1.1 g) was added. The resulting mixture was refluxed for 3 hours, then additional water (0.6 ml) was added and reflux continued for 5 hours. The organic solvent was then removed by evaporation, the residue was treated with saturated sodium bicarbonate solution (100 ml) and the resulting mixture was diluted with dilute hydrochloric acid to pH 6
Acidified. The mixture was extracted with ethyl acetate (2 × 100 ml) and the combined extracts were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was dried over phosphorus pentoxide in a vacuum desiccator overnight to give α-tert-butyl-N-trityl-L-4-methylglutamate (1.
06g) was obtained as a white foam.

α-tert-butyl-N-trityl-L-4-methylglutamate (0.76 g) and N-methylmorpholine (0.1
To a solution of 51 g) in dry tetrahydrofuran (3.5 ml) (cooled to −20 ° C.) was added, with stirring, isobutyl chloroformate (0.204 g). After 10 minutes di-tert-butyl-
L-glutamate hydrochloride (0.442 g) and N-methylmorpholine (0.151 g) in tetrahydrofuran (4.5 ml)
The mixture in was added. Stirring was continued at −20 ° C. for 10 minutes and then for 1.5 hours while allowing the mixture to warm to room temperature. The N-methylmorpholine hydrochloride was filtered off and the filtrate was concentrated in vacuo. The residue was purified by chromatography on a silica gel column using a gradient of ethyl acetate in distilled gasoline (7-20%) as eluent. The product solidified upon drying in vacuo over phosphorus pentoxide overnight.
Thus, tri-tert-butyl-N- [N- (trityl)
-L-γ-4-methylglutamyl] -L-glutamate (0.669 g) was obtained. 58-66 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 0.94 (d, J = 6.8 Hz, 3H, 4-
Meglu γ-CH ₃ ), 1.14 (s, 9H, 4-Meglu C (CH ₃ ) ₃ ),
1.39 (m, 19H, glu C (CH ₃ ) ₃ and 4-Meglu β-C
H), 1.85, 2.11 (2 xm, 3H, 4-Meglu β-CH and glu
β-CH ₂ ), 2.27 (t, J = 7.6 Hz, 2H, glu γ-CH ₂ ), 2.38
(M, 1H, 4-Meglu γ-CH), 2.68 (d, J = 9.0 Hz, 1H, 4-Me
glu NH), 3.04 (m, 1H, 4-Meglu α-CH), 4.13 (m, 1H, g
lu α-CH), 7.14-7.41 (m, 15H, TrH), 8.14 (d, J = 7.4
Hz, 1H, glu NH). Mass spectrum: (CI) m / e 701 (M + H) + Elemental analysis: Found C, 72.02; H, 8.02; N, 3.87% C 42 H 56 N 2 O 7 theory C, 71.97; H, 8.05; N Tri-tert-butyl-N- [N- (trityl) -L- in ethyl acetate (60 ml) containing 4.00% 10% Pd / C (0.160 g)
γ-4-methylglutamyl] -L-glutamate (0.60
The solution of 8 g) was stirred under hydrogen for 26 hours. Then additional catalyst (0.035 g) was added and stirring continued under hydrogen for 20 hours.
The catalyst was filtered off and the filtrate was concentrated in vacuo to give a white semi-solid mixture of triphenylmethane and tri-tert-butyl-L-γ-4-methylglutamyl-L-glutamate (0.450 g) I got

(3) Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-4-methylglutamyl-L-glutamic acid tri-tert in dry dimethylformamide (6.5 ml)
P-butyl-L-γ-4-methylglutamyl-L-glutamate (0.430 g) solution (cooled to 0 ° C.) under stirring
-[N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoic acid trifluoroacetate (0.327 g) was added, Then diethyl cyanophosphonate (0.231 g) and triethylamine (0.143 g) were added. The resulting solution was stirred at 0 ° C. in the dark for 30 minutes, then at room temperature for 1.5 hours. Then ethyl acetate (100 ml) and water (100
ml) was added to the reaction mixture. The two layers were separated and the aqueous layer was further extracted with ethyl acetate (2 × 100 ml). The combined ethyl acetate extracts were combined into a 10% aqueous citric acid solution (2
Times), washed with saturated sodium bicarbonate solution (150 ml), dilute aqueous sodium chloride solution (150 ml), water (100 ml), then dried over anhydrous sodium sulfate, filtered and freed of ethyl acetate in vacuo. The residue was purified by chromatography on a silica gel column using a gradient of methanol in ethyl acetate (0-2%) as eluent. The product is precipitated from dimethyl ether / petroleum ether at -20 DEG C. and thus tri-tert-butyl-Np- [N-
(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-4-methylglutamyl-L-
Glutamate (0.170 g) was obtained. 102-110 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.03 (d, J = 6.81 Hz, 3H, 4
−Meglu γ-CH ₃ ), 1.33,1.36,1.38 (3 xs, 27H, C (C
_{H 3) 3), 1.65,1.83,2.05 (} 3 xm, 4H, β-CH 2), 2.20
(T, J = 7.7 Hz, 2H, glu γ-CH ₂ ), 2.32 (s, 3 H, quinazoline 2-CH ₃ ), 2.50 (m, DMSO peak and 4-Meglu γ
−CH), 3.22 (s, 1H, C≡CH), 4.07 (m, 1H, glu α-CH),
4.21 (m, 1H, 4-Meglu α-CH), 4.34 (s, 2H, CH ₂ C≡
C), 4.78 (s, 2H , quinazoline _{6-CH 2), 6.82 (} d, J =
8.7Hz, 2H, 3 ', 5'-ArH), 7.54 (d, J = 8.4Hz, 1H, quinazoline 8-H), 7.68 (m, 3H, quinazoline 7-H and 2', 6'-ArH) , 7.96 (s, 1H, quinazoline 5-H), 8.
06 (d, J = 7.6Hz, 1H, glu NH), 8.20 (d, J = 7.7Hz, 1H, 4-
Meglu _L NH) 12.18 (s, 1H, lactam NH). Mass spectrum: (ESI) m / e 788 (M + H) + Elemental analysis: Found C, 64.91; H, 7.27; N, 8.79% C 43 H 57 N 5 O 9 · 0.5H 2 O Theoretical value C, 64.81; H, 7.33; N, 8.79% tri-tert-butyl-Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) ) Amino] benzoyl-L-γ-4-methylglutamyl-L-glutamate (0.071 g) was added to L-γ-glutamyl-N in Example 1 (3).
Treated with trifluoroacetic acid (6 ml) as described for -methyl-L-glutamate analog. Thus N
-P- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-
(Inyl) amino] benzoyl-L-γ-4-methylglutamyl-L-glutamic acid (containing 0.95 equivalents of trifluoroacetic acid, 0.4 equivalents of diethyl ether, and 1.2 equivalents of water; 0.053 g). 160-165 ° C (decomposition).

NMR spectrum: 1.02 (d, J = 6.8 Hz, 3H, 4-Meglu γ-C
_{H 3), 1.65,1.91,2.09 (3 xm} , 4H, β-CH 2), 2.25 (t, J
= 7.6Hz, 2H, glu γ-CH ₂ ), 2.39 (s, 3H, quinazoline 2
-CH _3), 2.50 (DMSO peaks and 4-Meglu γ-CH),
3.23 (s, 1H, C≡CH), 4.19 (m, 2H, glu α-CH and 4-
Muglu α-CH), 4.35 (s, 2H, CH ₂ C≡CH), 4.80 (s, 2H, quinazoline 6-CH ₂ ), 6.82 (d, J = 8.9 Hz, 2H, 3 ′, 5′-A
rH), 7.58 (d, J = 8, 4Hz, 1H, quinazoline 8-H), 7.7
4 (d, J = 8.7Hz, 3H, quinazoline 7-H and 2 ', 6'
-ArH), 7.99 (s, 1H, quinazoline 5-H), 8.10 (d, J
= 7.7Hz, 1H, glu NH), 8.26 (d, J = 7.7Hz, 1H, 4-Meglu N
H). Mass spectrum: (ESI) m / e 620 (M + H) + Elemental analysis: Found C, 53.05; H, 5.46; N, 8.97; F, 6.89% C 31 H 33 N 5 O 9 · 0.95CF 3 COOH · 0.4 Et ₂ O · 1.2H ₂ O theoretical value C, 53.18; H, 5.22; N, 8.99; F, 6.95% example 6: N-p- [N- ( 3,4- dihydro-2-methyl -
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl-L-γ-4,4-
Dimethylglutamyl-L-glutamic acid (1) α-tert-butyl-γ-allyl-N-trityl-
L-4,4-Dimethylglutamate To a suspension of L-glutamic acid (8.82 g) in dry allyl alcohol (300 ml) was added dropwise, with stirring, chlorotrimethylsilane (19 ml) under an argon atmosphere. The mixture was stirred at room temperature for 18 hours and then diethyl ether (2
Liter) resulted in the formation of a white precipitate. This was filtered off, washed further with diethyl ether and dried in vacuo over phosphorus pentoxide. Thus, γ-allyl acetate L
-Glutamate (10.1 g) was obtained as a white solid. Melting point 1
32-133 ° C (decomposition).

NMR spectrum: (CD ₃ SOCD ₃ ) 2.05 (m, 2H, β-CH ₂ ), 2.
54 (m, 2H, γ-CH ₂ ), 3.90 (t, J = 6.3 Hz, 1H, α-CH), 4.
55 (dm, J = 5.4 Hz, 2H, CH ₂ CH = CH ₂ ), 5.20 (dm, J = 9.3 Hz,
1H, CH = CH ₂ cis), 5.30 (dm, J = 16.3Hz, 1H, CH = CH 2 trans), 5.93 (m, 1H, CH = CH 2), 8.52 (bd s, 3H, NH 3). Mass spectrum: (CI) m / e 188 (M-Cl) γ-allyl-L-glutamate hydrochloride (8.94 g), acetic acid t
ert-butyl (250 ml) and 70% aqueous perchloric acid (6.
3g) was stirred at room temperature for 100 hours. The mixture was then cooled in an ice-water bath and extracted with cold 0.5N hydrochloric acid (3 × 80 ml). The combined aqueous extracts were immediately neutralized with solid sodium bicarbonate and then extracted with diethyl ether (3 × 120 ml). The combined extracts were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give α-tert-butyl-
γ-allyl-L-glutamate (10.0 g) was obtained as a colorless oil.

Α-tert-butyl-γ in dry chloroform (35 ml)
To a solution of -allyl-L-glutamate (8.4 g) was added, with stirring, triethylamine (5.25 g), followed by a solution of trityl chloride (7.23 g) in chloroform (15 ml), followed by lead nitrate (4.3 g). Was added. After stirring at room temperature for 17 hours, a white solid formed. The solid was filtered off, the filtrate was concentrated in vacuo, the residue was treated with hexane (40ml) and left in the refrigerator overnight. The hexane solution is then filtered, the filtrate is concentrated in vacuo and the residue is concentrated on a silica gel column with a gradient of ethyl acetate in distilled petroleum ether (4-
(6%) was purified by chromatography using as eluent. Thus, α-tert-butyl-γ-allyl-N
-Trityl-L-glutamate (8.10 g) was obtained as a yellow oil.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.13 (s, 9H, C (CH ₃ ) ₃ , 1.
80,1.90 (2 xm, 2H, β-CH ₂ ) 2.20,2.46 (2 xm, 2H, γ
−CH ₂ ), 2.80 (d, J = 9.0 Hz, 1H, NH), 3.17 (m, 1H, α-C
H), 4.54 (d, J = 5.5Hz, 2H, CO 2 CH 2 CH = CH 2), 5.20 (dm, J
= 10.4Hz, 1H, CH = CH ₂ cis), 5.29 (dm, J = 15.8Hz, 1H, CH
= CH ₂ trans), 6.88 (m, 1H, CH = CH 2), 7.15-7.40
(M, 15H, TrH). Mass spectrum: (CI) m / e 486 (M + H) + Elemental analysis: Found C, 76.46; H, 7.39; N, 2.82% C 31 H 35 NO 4 theory C, 76.67; H, 7.26; N, 2.88 % To a solution of potassium hexamethyldisilazide (23.68 ml, 0.5 M in toluene) in dry tetrahydrofuran (22 ml) was added α-tert-butyl in dry tetrahydrofuran (26 ml) while stirring at −78 ° C. under argon. A solution of -γ-allyl-N-trityl-L-glutamate (3.59 g) was added over 5 minutes. The resulting yellow solution was -78
Stirred at C for 55 min, then added methyl iodide (2.10 g). Stirring was continued at −78 ° C. for 30 minutes, then the white slurry was poured into a saturated ammonium chloride solution (200 ml). The resulting mixture was diluted with diethyl ether (3 in 150 ml).
Times), the combined ether extracts were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography on a silica gel column using a gradient of ethyl acetate in petroleum ether (6-8%) as eluent. Thus, α-tert-butyl-γ-
Allyl-N-trityl-L-4-methylglutamate (3.26 g) was obtained as an oil. It was a mixture of the two diastereoisomers (2S, 4S) and (2S, 4R).

NMR spectrum: (CD ₃ SOCD ₃ ) 1.00,1.07 (2 xd, J = 7.0
Hz, 3H, γ-CH ₃ ), 1.12, 1.15 (2 xs, 9H, C (CH ₃ ) ₃ ),
1.38,1.57 (2 xm, 1H, β-CH), 1.95,2.15 (2 xm, 1H,
β-CH), 2.45 (m, 1H, γ-CH), 2.80 (t, J = 9.4Hz, 1H, N
H), 3.12 (m, 1H, α-CH), 4.43 (ABdd, J ₁ = 13.7 Hz, J ₂ =
_{5.4Hz, 1H, CH 2 CH =} CH 2), 4.60 (d, J = 5.3Hz, 1H, CH 2 CH = C
_{H 2), 5.10-5.35 (m,} 2H, CH = CH 2), 5.70-6.00 (m, 1H, C
_{H = CH 2), 7.16-7.40 (} m, 15H, TrH). Mass spectrum: (CI) m / e 500 (M + H) + Elemental analysis: Found C, 75.74; H, 7.47; N, 2.73% C 32 H 37 NO 4 · 0.4H 2 O Theoretical value C, 75.83; H, 7.52; N, 2.76% A solution of potassium hexamethyldisilazide (16.2 ml, 0.5M in toluene) in dry tetrahydrofuran (16 ml) was stirred at -78 ° C under argon with stirring in dry tetrahydrofuran (16 ml). α-tert-butyl-γ-allyl-N-trityl-L-4-methylglutamate (2.24
The solution of g) was added over 5 minutes. The resulting solution was stirred at -78 ° C for 55 minutes, then methyl iodide (1.12m
l) was added. Stirring was continued at -78 ° C for 2 hours, then the yellow slurry was added to a saturated ammonium chloride solution (200 ml).
Was injected. The aqueous solution was diluted with diethyl ether (150 ml
Times), the combined ether extracts were dried over anhydrous sodium sulfate, filtered, and the solution was concentrated in vacuo.
The residue was purified by chromatography on a silica gel column using a gradient of ethyl acetate in distilled petroleum ether (3-6%) as eluent. Thus, α-tert-butyl-γ-allyl-N-trityl-L-4,4-dimethylglutamate (2.08 g) was obtained as a colorless oil.

NMR spectrum: (CD ₃ SOCD ₃ ) 0.94, 0.96 (2 xs, 6H, γ
_{-CH 3), 1.20 (s,} 9H, C (CH 3) 3), 1.34 (dd, J 1 = 14.2H
_{z, J 2 = 3.7Hz, 1H} , β-CH), 2.06 (dd, J 1 = 14.3Hz, J 2 = 8.
6Hz, 1H, β-CH), 2.78 (d, J = 8.9Hz, 1H, NH), 3.09 (m, 1
H, α-CH), 4.50 (ABddd, J ₁ = 15.0 Hz, J ₂ = 5.0 Hz, J ₃ = 1.
_{2Hz, 2H, CH 2 CH =} CH 2), 5.16-5.30 (m, 2H, CH = CH 2), 5.8
7 (m, 1H, CH = CH 2), 7.16-7.40 (m, 15H, TrH). Mass spectrum: (CI) m / e 514 (M + H) + Elemental analysis: Found C, 75.96; H, 7.61; N, 2.66% C 33 H 39 NO 4 · 0.5H 2 O Theoretical value C, 75.83; H, 7.71; N, 2.68% (2) tri-tert-butyl-L-γ-4,4-dimethylglutamyl-L-glutamate α-tert-butyl- in dry dichloromethane (11 ml)
While stirring under argon with a solution of γ-allyl-N-trityl-L-4,4-dimethylglutamate (1.4 g),
Tetrakis (triphenylphosphine) palladium (0) (0.091 g) was added, followed by pyrrolidine (0.35 m
l) was added. The resulting yellow solution was stirred at room temperature for 20 minutes, then diethyl ether (100 ml) and 1N hydrochloric acid (50 ml) were added. The two layers were separated and the aqueous layer was further washed with diethyl ether (100ml). The combined ether extracts were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was treated with diethyl ether (30 ml), the yellow precipitate was filtered off and the filtrate was concentrated in vacuo to give α
-Tert-Butyl-N-trityl-L-4,4-dimethylglutamate (1.25 g) was obtained as a white foam.

α-tert-butyl-N-trityl-L-4,4-dimethylglutamate (0.710 g) and (1H-1,2,3-benzotriazol-1-yloxy) -tris (pyrrolidino) -phosphonium hexafluorophosphate ( PyBO
To a solution of P) (0.935 g) in dry dichloromethane (1.9 ml) was added, with stirring, diisopropylethylammine (0.464 g), followed by 4-dimethylaminopyridine (0.109 g) and di-tert-butyl- L-glutamate hydrochloride (0.531 g) was added. After stirring at room temperature for 3 hours, the solvent was removed by evaporation and the yellow residue was purified on a silica gel column with a gradient of ethyl acetate in hexane (5-ethyl acetate).
(20%) as the eluent. Thus, tri-tert-butyl-N- [N-
(Trityl-L-γ-4,4-dimethylglutamyl) -L
-Glutamate (0.445 g) was obtained as an oil.

NMR Spectrum: 0.80,0.95 (2 xs, 6H, 2 x γ-CH 3), 1.
19 (s, 9H, 4-diMeglu C (CH ₃ ) ₃ ), 1.34 (s, 9H, glu C
(CH ₃ ) ₃ ), 1.39 (m, 10H, glu C (CH ₃ ) ₃ and 4-di)
Meglu β-CH), 1.78-1.98 (m, 3H, glu β-CH ₂ and 4-diMeglu β-CH), 2.22 (t, J = 7.2 Hz, 2H, glu γ-C
H ₂ ), 2.80 (d, J = 8.2 Hz, 1H, 4-diMeglu NH), 2.98 (m, 1
H, 4-diMeglu α-CH), 4.04 (m, 1H, glu α-CH), 7.14
−7.40 (m, 15H, TrH). Mass spectrum: (CI) m / e 715 (M + H) + Elemental analysis: Found C, 71.65; H, 8.22; N, 3.84% C 34 H 58 N 2 O 7 · 0.3H 2 O Theoretical value C, 71.70; H, 8.20; N, 3.89% Tri-tert-butyl-N- [N- (trityl-L-γ) in ethyl acetate (50 ml) containing 10% Pd / C (0.065 g).
-4,4-dimethylglutamyl] -L-glutamate (0.4
30g) was stirred under hydrogen for 26 hours. Then additional catalyst (0.075 g) was added and stirring continued under hydrogen for 16 hours.
The catalyst was then filtered off and the filtrate was concentrated in vacuo to give a white semi-solid, triphenylmethane and tri-tert-
A mixture of butyl-L-γ-4,4-dimethylglutamyl-L-glutamate (0.400 g) was obtained.

(3) Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl] -L-γ-4,4
-Dimethylglutamyl-L-glutamic acid tri-tert in dry dimethylformamide (7.0 ml)
-Butyl-L-γ-4,4-dimethylglutamyl-L-glutamate (0.360 g) in solution (cooled to 0 ° C under argon) under stirring with p- [N- (3,4-dihydro-2). -Methyl-4-oxoquinazolin-6-ylmethyl) -N-
(Prop-2-ynyl) amino] benzoic acid trifluoroacetate (0.200 g) was added, followed by diethyl cyanophosphonate (0.154 g) and triethylamine (0.095 g). The resulting solution was allowed to stand at 0 ° C. in the dark for 10 minutes,
Then, the mixture was stirred at room temperature for 1.5 hours. Then ethyl acetate (70m
l) and water (70 ml) were added to the reaction mixture. The two layers were separated and the aqueous layer was further extracted with ethyl acetate (2x50ml). The combined ethyl acetate extracts were washed with 10% aqueous citric acid (100 ml), saturated aqueous sodium bicarbonate (100 ml), dilute aqueous sodium chloride (100 ml), and water (100 ml), then dried over anhydrous sodium sulfate, Filtered,
Concentrated in vacuo. The residue was purified by chromatography on a silica gel column using a gradient of methanol in ethyl acetate (0-2%) as eluent. The product is precipitated from dimethyl ether / petroleum ether at −20 ° C. and tri-tert-butyl-Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N -(Prop-2-ynyl) amino] benzoyl-L-γ-4,4-dimethylglutamyl-L-glutamate (0.200 g) was obtained as a white powder. Melting point 101-104
° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.11,1.16 (2 × s, 6H, 4-
DiMeglu γ-CH ₃ ), 1.30, 1.36 (2 xs, 27H, C (C
_{H 3) 3), 1.95 (} m, 4H, β-CH 2), 2.21 (t, J = 7.7Hz, 2H,
glu γ-CH ₂ ), 2.32 (s, 3H, quinazoline 2-CH ₃ ), 3.2
2 (s, 1H, C≡CH), 4.08 (m, 1H, glu α-CH), 4.23 (m, 1
H, 4-diMeglu α-CH), 4.31 (s, 2H, CH ₂ C≡C), 4.76
(S, 2H, quinazoline _{6-CH 2), 6.81 (} d, J = 8.7z, 2H,
3 ', 5'-ArH), 7.52 (d, J = 8.4Hz, 1H, quinazoline 8
-H), 7.66 (m, 4H, quinazoline 7-H, 2 ', 6'-ArH and 4-diMeglu NH), 7.95 (s, 1H, quinazoline 5-
H), 8.19 (d, J = 7.3 Hz, 1H, glu NH), 12.19 (s, 1H, lactam NH). Mass spectrum: (ESI) m / e 801 (M ⁺ ) Elemental analysis: found C, 66.03; H, 7.68; N, 8.50% C ₄₄ H ₅₉ N ₅ O ₉ theoretical C, 65.90; H, 7.41; N , 8.73% tri-tert-butyl-Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoyl- L-γ-4,4-dimethylglutamyl-L-glutamate (0.068 g) was added to L-γ-glutamyl-in Example 1 (3).
Treated with trifluoroacetic acid (7 ml) as described for the N-methyl-L-glutamate analog. Thus Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2
-Inyl) amino] benzoyl-L-γ-4,4-dimethylglutamyl-L-glutamic acid (containing 0.6 equivalents of trifluoroacetic acid and 0.8 equivalents of water; 0.050 g). Melting point> 140 ° C (decomposition).

NMR spectrum (CD ₃ SOCD ₃ ): 1.12, 1.17 (2 xs, 6H, 4-
Di-Meglu γ-CH ₃ ), 1.84-2.13 (m, 4H, β-CH ₂ ), 2.25
(T, J = 7.4Hz, 2H, glu γ-CH ₂ ), 2.38 (s, 3H, quinazoline 2-CH ₃ ), 3.23 (s, 1H, C≡CH), 4.20 (m, 1H, glu α
-CH), 4.33 (m, 3H , CH 2 C≡CH and 4-di Meglu α-C
H), 4.79 (s, 2H , quinazoline _{6-CH 2), 6.82 (} d, J = 8.
6Hz, 2H, 3 ', 5'-ArH), 7.57 (zd, J = 8.4Hz, 1H, quinazoline 8-H), 7.71 (m, 4H, quinazoline 7-H, 2',
6'-ArH and 4-diMeglu NH), 7.99 (s, 1H, quinazoline 5-H), 8.22 (d, J = 7.30 Hz, 1H, glu NH). Mass spectrum: (ESI) m / e 634 (M + H) + Elemental analysis: Found C, 55.71; H, 5.41; N, 9.60; F, 4.64% C 32 H 35 N 5 O 9 · 0.6THF · 0.8H 2 O Theoretical C, 55.66; H, 5.23; N, 9.77; F, 4.77% Example 7: Np- [N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazoline-6 Ylmethyl) -N-
(Prop-2-ynyl) amino] benzoyl-L-γ-
Glutamyl-N-methyl-L-glutamic acid The method described in Example 1 (3) was converted to Np- [N-
Instead of (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoic acid trifluoroacetate, Np-
[N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoic acid trifluoroacetate (0.475 g; UK Patent No. (Prepared as described in Example 10 of 2 202 847B) as starting material. Thus, tri-tert-butyl-Np- [N- (3,4-dihydro-2,7
-Dimethyl-4-oxoquinazolin-6-ylmethyl)
-N- (prop-2-ynyl) amino] benzoyl-L
-Γ-glutamyl-N-methyl-L-glutamate (0.
340 g). 114-116 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.36 (m, 27H, C (C
_{H 3) 3), 1.74-2.20 (} m) and 2.42 (m 'concealment) (8
H, β-CH ₂ and γ-CH ₂ ), 2.31 (s, 3H, quinazoline 2
_{-CH 3), 2.44 (s,} 3H, quinazoline 7-CH _3), 2.62,2.82
(2 xs, 3H, N- CH 3), 3.15 (s, 1H, C≡CH), 4.26 (S, 2H,
CH ₂ C≡C), 4.30 (m concealed, 1H, glu α-CH), 4.44, 4.7
_{9 (2 x dd, J 1} = 10.58Hz, J2 = 4.86Hz, 1H, N-Me glu α-
CH), 4.66 (s, 2H , quinazoline _{6-CH 2), 6.81 (} d, J =
8.95Hz, 2H, 3 ', 5'-ArH), 7.43 (s, 1H, quinazoline 8
-H), 7.74 (d, J = 7.74 Hz, 3H, quinazoline 5-H and 2 ', 6'-ArH), 8.22 (t, J = 7.44 Hz, 1H, glu NH), 1
2.00 (s, 1H, lactam NH). Mass spectrum: (ESI) m / e 802 (M + H) + Elemental analysis: Found C, 65.37; H, 7.35; N, 8.63% C 44 H 59 N 5 O 9 · 0.25H 2 O Theoretical value C, 65.53; H, 7.43; N, 8.68% tri-tert-butyl-Np- [N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2 A solution of -inyl) amino] benzoyl-L-γ-glutamyl-N-methyl-L-glutamate (0.200 g) in trifluoroacetic acid (15 ml) was stirred at room temperature in the dark for 1.2 hours. The solution was then concentrated in vacuo and the residue was triturated with dry diethyl ether (20ml). The white solid was isolated by filtration, washed with diethyl ether and dried in vacuum.
Thus Np- [N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N-
(Prop-2-ynyl) amino] benzoyl-L-γ-
Glutamyl-N-methyl-L-glutamic acid trifluoroacetate (0.180 g) was obtained. Decomposes at 170 ° C.

NMR _{Spectrum: (CD 3 SOCD 3) 1.82-2.23} (m) and
2.46 (m concealment) (8H, β-CH ₂ and γ-CH _2), 2.23,
2.48 (2 xs, 6H, quinazoline 2-CH ₃ and quinazoline 7-CH ₃ ), 2.65, 2.81 (2 xs, 3H, N-CH ₃ ), 3.23 (s, 1
H, C≡CH), 4.31 (m, 3H, CH ₂ C≡C and glu α-CH), 4.
_{56,4.91 (2 x dd, J 1} = 10.87Hz, J 2 = 4.38Hz, 1H, N-Me gl
u α-CH), 4.71 (s, 2H, quinazoline 6-CH ₂ ), 6.79
(D, J = 8.89 Hz, 2H, 3 ', 5'-ArH), 7.48 (s, 1H, quinazoline 8-H), 7.76 (m, 3H, 2', 6'-ArH and quinazoline 5-H) , 8.33 (m, 1H, glu NH). Mass spectrum: (ESI) m / e 634 (M + H) + Elemental analysis: Found C, 53.23; H, 5.00; N, 8.86; F, 7.33% C 32 H 35 N 5 O 9 · 1CF 3 COOH · 1H 2 O Theoretical C, 53.33; H, 5.00; N, 9.15; F, 7.44% Example 8: Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] -o-fluorobenzoyl-
L-γ-glutamyl-N-methyl-L-glutamic acid The method described in Example 1 (3) was converted to Np- [N-
Instead of (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoic acid trifluoroacetate, Np-
[N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] -o-fluorobenzoic acid trifluoroacetate (0.479 g; UK) (Prepared as described in Example 6 of WO-A-2 253 849A) as starting material. Thus, tri-tert-butyl-Np- [N-
(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] -o-fluorobenzoyl-L-γ-glutamyl-
N-methyl-L-glutamate (0.430 g) was obtained. Melting point
110-112 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.36 (m, 27H, C (C
_{H 3) 3), 1.80,2.20,2.42 (} 2 xm, 8H, β-CH 2 , and γ
_{-CH 2), 2.33 (s,} 3H, quinazoline 2-CH _3), 2.60,2.80
(2 xs, 3H, N- CH 3), 3.27 (s, 1H, C≡CH), 4.29 (m, 1H,
glu α-CH), 4.37 (s, 2H, CH ₂ C≡C), 4.50 (dd) and 4.81 (dd concealment), (1H, J ₁ = 10.15 Hz, J ₂ = 4.78 Hz, N
-Me glu α-CH), 4.79 (s, 2H, quinazoline 6-C
_{H 2), 6.61 (d,} J = 14.56Hz, 1H, 3'-ArH), 6.66 (d, J =
8.63Hz, 1H, 5'-ArH) , 7.52 (m, 2H, 6'-ArH and quinazoline 8-H), 7.68 (dd , J 1 = 8.41Hz, J 2 = 1.81Hz, 1
H, quinazoline 7-H), 7.95 (s, 1H, quinazoline 5-
H), 8.02 (dd concealment) and 8.07 (dd) (1H, J 1 = 7.04
_{Hz, J 2 = 4.2Hz, glu} NH), 12.21 (s, 1H, Ramutamu NH). Mass spectrum: (ESI) m / e 806 (M + H) + Elemental analysis: Found C, 63.68; H, 6.97; N, 8.62; F, 2.54% C 43 H 56 FN 5 O 9 · 0.25H 2 O Theoretical value C, 63.73; H, 7.03; N, 8.64; F, 2.34% tri-tert-butyl-Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)- N- (prop-2-ynyl) amino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl-L
Glutamate (0.220 g) in trifluoroacetic acid (15 m
The solution in l) was stirred at room temperature in the dark for 1.2 hours. The solution was then concentrated in vacuo and the residue was triturated with dry diethyl ether (20ml). The white solid was isolated by filtration, washed with diethyl ether and dried in vacuum. Thus Np- [N- (3,4-dihydro-
2-methyl-4-oxoquinazolin-6-ylmethyl)
-N- (prop-2-ynyl) amino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid (0.213 g) was obtained. Melting point: 120 ° C, decomposed.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.82-2.20 (m) and
2.41 (m concealed) (8H, β-CH ₂ and γ-CH ₂ ), 2.43
(S, 3H, quinazoline _{2-CH 3), 2.63,2.80 (} 2 xs, 3H, N
−CH ₃ ), 3.27 (s, 1H, C≡CH), 4.37 (m, 3H, CH ₂ C≡C and glu α-CH), 4.54, 4.90 (2 x dd, J ₁ = 11.23 Hz, J ₂ =
4.35Hz, 1H, N-Me glu α-CH), 4.82 (s, 2H, quinazoline _{6-CH 2), 6.61 (} d, J = 15.01Hz, 1H, 3'-ArH), 6.66
(D, J = 8.10 Hz, 1H, 5'-ArH), 7.52 (t, J = 8.73 Hz, 1H,
6'-ArH), 7.61 (d, J = 8.42Hz, 1H, quinazoline 8-
H), 7.77 (d, J = 8.48 Hz, 1H, quinazoline 7-H), 8.0
0 (s, 1H, quinazoline 5-H), 8.05 (m, 1H, glu NH). Mass spectrum: (ESI) m / e 638 (M + H) + Elemental analysis: Found C, 50.45; H, 4.56; N, 8.22; F, 11.12% C 31 H 32 FN 5 O 9 · 1.3CF 3 COOH · 1H ₂ O · 1Et ₂ O theoretical value C, 50.47; H, 4.59; N, 8.55; F, 11.37% example 9: N-p- [N- ( 3,4- dihydro-2,7-dimethyl-4- Oxoquinazolin-6-ylmethyl) -N-
(Prop-2-ynyl) amino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid The method described in Example 1 (3) was converted to Np- [N-
Instead of (3,4-dihydro-2-methyl-4-oxaquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoic acid trifluoroacetate, Np-
[N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] -o-fluorobenzoic acid trifluoroacetate (0.494 g) Prepared as described in Example 7 of GB-A-2 253 849A) as starting material. Thus, tri-tert-butyl-Np- [N-
(3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl- L-glutamate (0.400 g) was obtained. 122-124 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.35 (m, 27H, C (C
_{H 3) 3), 1.70-2.20 (} m) and 2.40 (m concealment) (8
H, β-CH ₂ and γ-CH ₂ ), 2.30 (s, 3H, quinazoline 2
_{-CH 3), 2.43 (s,} 3H, quinazoline 7-CH _3), 2.60,2.80
(2 xs, 3H, N- CH 3), 3.25 (s, 1H, C≡CH), 4.30 (m, 3H,
CH ₂ C≡C and glu α-CH), 4.50, 4.79 (2 × dd, J ₁ = 1
_{0.68Hz, J 2 = 4.61Hz, 1H} , N-Me glu α-CH), 4.68 (s, 2
H, quinazoline _{6-CH 2), 6.61 (} m, 2H, 3 ', 5'-Ar
H), 7.44 (s, 1H, quinazoline 8-H), 7.51 (t, J = 8.6
7Hz, 1H, 6'-ArH), 7.68 (s, 1H, quinazoline 5-H),
_{8.00,8.08 (2 x dd, J 1} = 6.98Hz, J 2 = 4.30Hz, 1H, glu N
H), 12.10 (s, 1H, lactam NH). Mass spectrum: (ESI) m / e 820 (M + H) ⁺ Elemental analysis: Found C, 64.25; H, 7.10; N, 8.51; F, 2.36% C ₄₄ H ₅₈ FN ₅ O ₉ Theoretical C, 64.45; H , 7.13; N, 8.5
4; F, 2.32% tri-tert-butyl-N-p- [N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl ) Amino] -o-
A solution of fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamate (0.215 g) in trifluoroacetic acid (16 ml) was stirred at room temperature in the dark for 1.2 hours. The solution was then concentrated in vacuo and the residue was triturated with dry diethyl ether (20ml). The white solid was isolated by filtration, washed with diethyl ether and dried in vacuum. Thus Np- [N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] -o
-Fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid (0.205 g) was obtained. Melting point 150 ° C,
Disassembly.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.82-2.20 (m) and
2.46 (m concealment) (8H, β-CH ₂ and γ-CH _2), 2.45,
2.48 (2 xs, 6H, quinazoline 2-CH ₃ and quinazoline _{7-CH 3), 2.63,2.81 (} 2 xs, 3H, N-CH 3), 3.25 (s, 1
H, C≡CH), 4.33 (m, 3H, CH ₂ C≡C and glu α-CH), 4.
_{58,4.90 (2 x dd, J 1} = 10.94Hz, J 2 = 4.43Hz, 1H, N-Me gl
u α-CH), 4.74 (s, 2H, quinazoline 6-CH ₂ ), 6.61
(M, 2H, 3 ', 5'-ArH), 7.49 (s, 1H, quinazoline 8-
H), 7.54 (t, J = 8.73 Hz, 1H, 6'-ArH), 7.73 (s, 1H, quinazoline 5-H), 8.08 (m, 1H, glu NH). Mass spectrum: (ESI) m / e 652 (M + H) + Elemental analysis: Found C, 50.90; H, 4.52; N, 8.40; F, 11.58% C 32 H 34 FN 5 O 9 · 1.3CF 3 COOH · 1H ₂ O theoretical value C, 50.81; H, 4.60; N, 8.56; F, 11.38% example 10: N-p- [N- ( 3,4- dihydro-2,7-dimethyl-4-oxoquinazolin -6 -Ylmethyl) -N-methylamino] benzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid The method described in Example 1 (3) was replaced with Np- [N-
Instead of (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoic acid trifluoroacetate, Np-
[N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino] benzoic acid trifluoroacetate (0.450 g; UK Patent Application Publication No. 2 253 849A) Prepared as described in Example 11 of the specification). Thus, tri-tert-
Butyl-Np- [N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino] benzoyl-L-γ-glutamyl-N-methyl-L -Glutamate (0.260 g) was obtained. Melting point 139-1
40 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.35 (m, 27H, C (C
_{H 3) 3), 1.80-2.18 (} m) and 2.40 (m concealment) (8
H, β-CH ₂ and γ-CH ₂ ), 2.30 (s, 3H, quinazoline 2
_{-CH 3), 2.43 (s,} 3H, quinazoline 7-CH _3), 2.61,2.81
(2 xs, 3H, N- CH 3), 3.13 (s, 3H, 10-N-CH 3), 4.25
(M, 1H, glu α-CH), 4.69 (s, 2H, quinazoline 6-C
_{H 2), 4.48,4.80 (2 x} dd, J 1 = 10.72Hz, J 2 = 4.66Hz, 1H, N
−Me glu α-CH), 6.70 (d, J = 8.88 Hz, 2H, 3 ′, 5′-Ar
H), 7.44, 7.53 (2 xs, 2H, quinazoline 5-H and quinazoline 8-H), 7.71 (d, J = 8.77 Hz, 2H, 2 ', 6'-A
rH), 8.24 (t, J = 7.51 Hz, 1H, glu NH), 12.10 (s, 1H, lamtam NH). Mass spectrum: (ESI) m / e 778 (M + H) + Elemental analysis: Found C, 64.32; H, 7.58; N, 8.92% C 42 H 59 N 5 O 9 · 0.3H 2 O Theoretical value C, 64.40; H, 7.67; N, 8.94% tri-tert-butyl-Np- [N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino] benzoyl -L-γ-glutamyl-N-methyl-L-glutamate (0.135 g)
At room temperature in the dark in trifluoroacetic acid (12 ml)
Stir for 1.2 hours. The solution is then concentrated in vacuo,
The residue was triturated with dry diethyl ether and dried in vacuo. Thus Np- [N- (3,4-dihydro-
2,7-Dimethyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino] benzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid (0.120 g) was obtained. 155 ° C, decomposes.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.80-2.16 (m) and
2.41 (m concealed) (8H, β-CH ₂ and γ-CH ₂ ), 2.40,
2.47 (2 xs, 6H, quinazoline 2-CH ₃ and quinazoline _{7-CH 3), 2.64,2.80 (} 2 xs, 3H, N-CH 3), 3.13 (s, 3
_{H, 10-N-CH 3} ), 4.31 (m, 1H, glu α-CH), 4.57,4.91
_{(2 x dd, J 1 =} 11.0Hz, J 2 = 4.31Hz, 1H, N-Me glu α-C
H), 4.72 (s, 2H , quinazoline _{6-CH 2), 6.71 (} d, J = 8.
71 Hz, 2H, 3 ', 5'-ArH), 7.47, 7.53 (2 xs, 2H, quinazoline 5-H and quinazoline 8-H), 7.73 (d, J =
8.53Hz, 2H, 2 ', 6'-ArH), 8.28 (d, J = 7.18Hz, H, glu N
H). Mass spectrum: (ESI) m / e 610 (M + H) + Elemental analysis: Found C, 51.29; H, 5.14; N, 9.00; F, 7.73% C 30 H 35 N 5 O 9 · 1CF 3 COOH · 1.5H ₂ O theoretical value C, 51.20; H, 5.24; N, 9.33; F, 7.59% example 11: N-p- [N- ( 3,4- dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl ) -N-Methylamino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid The method described in Example 1 (3) was replaced with Np- [N-
Instead of (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoic acid trifluoroacetate, Np-
[N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino] -o-
Repeated with fluorobenzoic acid trifluoroacetate (0.410 g; prepared as described in Example 12 of GB 2 253 849A). Thus, tri-tert-butyl-Np- [N- (3,4-dihydro-2-methyl-
4-oxoquinazolin-6-ylmethyl) -N-methylamino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamate (0.465 g) was obtained.
107-109 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.35 (m, 27H, C (C
_{H 3) 3), 1.80-2.18,2.42 (} 2 xm, 8H, β-CH 2 , and γ
_{-CH 2), 2.32 (s,} 3H, quinazoline 2-CH _3), 2.60,2.80
(2 xs, 3H, N- CH 3), 3.13 (s, 3H, 10-N-CH 3), 4.29
(M, 1H, glu α-CH), 4.49 (m) and 4.79 (m concealed) (1H, N-Me glu α-CH), 4.78 (s, 2H, quinazoline 6-CH ₂ ), 6.54 (d, J = 16.11Hz, 1H, 3'-ArH), 6.61
(D, 9.33Hz, 1H, 5'-ArH), 7.56 (m, 3H, 6'-ArH, quinazoline 7-H and quinazoline 8-H), 7.84
(S, 1H, quinazoline 5-H), 7.87, 7.97 (2 xt, J = 6.
20Hz, 1H, glu NH), 12.22 (s, 1H, lactam NH). Mass spectrum: (ESI) m / e 782 (M + H) + Elemental analysis: Found C, 62.70; H, 7.21; N, 9.02; F, 2.62% C 41 H 56 N 5 O 9 F theory C, 62.98; H, 7.22; N, 8.96; F, 2.43% tri-tert-butyl-Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino ] -O-fluorobenzoyl-
A solution of L-γ-glutamyl-N-methyl-L-glutamate (0.295 g) in trifluoroacetic acid (23 ml) was stirred at room temperature in the dark for 1.2 hours. The solution was then concentrated in vacuo, the residue was triturated with dry diethyl ether and dried in vacuum. Thus Np-
[N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino] -o-
Fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid (0.270 g) was obtained. Melting point 110 ° C, decomposition.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.82-2.18 (m) and
2.40 (m concealed) (8H, β-CH ₂ and γ-CH ₂ ), 2.45
(S, 3H, quinazoline _{2-CH 3), 2.63,2.81 (} 2 xs, 3H, N
−CH ₃ ), 3.13 (s, 3H, 10-N-CH ₃ ), 4.34 (m, 1H, glu α
−CH), 4.53, 4.90 (2 x dd, J ₁ = 10.91 Hz, J ₂ = 4.36 Hz, 1
H, N-Me glu α-CH), 4.83 (s, 2H, quinazoline 6-CH
₂ ), 6.56 (d, J = 15.86 Hz, 1H, 3'-ArH), 6.62 (d, J = 9.
1Hz, 5'-ArH), 7.53 (t, J = 9.02Hz, 1H, 6'-ArH), 7.6
2 (d, J = 8.43 Hz, 1H, quinazoline 8-H), 7.70 (dd, J _{1
= 8.43Hz, J 2 = 1.82Hz,} 1H, quinazoline 7-H), 7.89
(D, J = 1.52 Hz, 1H, quinazoline 5-H), 7.97 (t, J =
5.50Hz, glu NH). Mass spectrum: (ESI) m / e 614 (M + H) + Elemental analysis: Found C, 48.32; H, 4.72; N, 8.12; F, 13.16% C 29 H 32 FN 5 O 9 · 1.6CF 3 COOH · 0.4 Et ₂ O · H ₂ O theoretical value C, 48.12; H, 4.73; N, 8.30; F, 13.05% example 12: N-p- [N- ( 3,4- dihydro-2,7-dimethyl-4 -Oxoquinazolin-6-ylmethyl) -N-methylamino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid The method described in Example 1 (3) was replaced with Np- [ N-
Instead of (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2-ynyl) amino] benzoic acid trifluoroacetate, Np-
[N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino] -o
Repeated using -fluorobenzoic acid trifluoroacetate (0.478 g; prepared as described in Example 13 of GB-A-2 253 849A). Thus, tri-tert-
Butyl-Np- [N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino] -o-fluorobenzoyl-L-γ-glutamyl-N -Methyl-L-glutamate (0.518 g) was obtained. 162-164 ° C.

NMR spectrum: (CD ₃ SOCD ₃ ) 1.36 (m, 27H, C (C
H ₃ ) ₃ ), 1.73-2.18 (m) and 2.40 (m concealed) (8
H, β-CH ₂ and γ-CH ₂ ), 2.30 (s, 3H, quinazoline 2
_{-CH 3), 2.42 (s,} 3H, quinazoline 7-CH _3), 2.60,2.80
(2 xs, 3H, N- CH 3), 3.11 (s, 3H, 10-N-CH 3), 4.29
(M, 1H, gu α-CH), 4.50, 4.79 (2 x dd, J ₁ = 10.72 Hz, J
_{2 = 4.53Hz, 1H, N-} Me glu α-CH), 4.69 (s, 2H, quinazoline _{6-CH 2), 6.51 (} d, J = 14.38Hz, 1H, 3'-ArH),
6.56 (d, J = 8.10 Hz, 1H, 5'-ArH), 7.48 (m, 3H, 6'-Ar
H, quinazoline 5-H and quinazoline 8-H), 7.8
8,7.98 (2 xt, J = 6.13 Hz, 1H, glu NH), 12.12 (s, 1H, lamtam NH). Mass spectrum: (ESI) m / e 796 (M + H) + Elemental analysis: Found C, 63.19; H, 7.31; N, 8.80; F, 2.44% C 42 H 58 N 5 O 9 F theory C, 63.38; H, 7.34; N, 8.80; F, 2.39% tri-tert-butyl-Np- [N- (3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N- Methylamino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl-L-glutamate (0.296 g) was treated in trifluoroacetic acid (23 ml),
Stirred at room temperature in the dark for 1.2 hours. The solution was then concentrated in vacuo, the residue was triturated with dry diethyl ether and dried in vacuum. Thus, Np- [N-
(3,4-dihydro-2,7-dimethyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino] -o-fluorobenzoyl-L-γ-glutamyl-N-methyl-L
-Glutamic acid (0.285 g) was obtained. 155 ° C, decomposes.

NMR _{Spectrum: (CD 3 SOCD 3) 1.80-2.18} (m) and
2.40 (m concealment) (8H, β-CH ₂ and γ-CH _2), 2.43,
2.47 (2 xs, 6H, quinazoline 2-CH ₃ and quinazoline _{7-CH 3), 2.63,2.81 (} 2 xs, 3H, N-CH 3), 3.13 (s, 3
H, 10-N-CH ₃ ), 4.34 (m, 1H, glu _L α-CH), 4.54,4.90
_{(2 x dd, J 1 =} 10.98Hz, J 2 = 4.43Hz, 1H, N-Me glu α-C
H), 4.73 (s, 2H , quinazoline _{6-CH 2), 6.53 (} d, J = 1
3.10Hz, 1H, 3'-ArH), 6.57 (d, J = 6.90Hz, 1H, 5'-Ar
H), 7.48, 7.50 (2 xs, 2H, quinazoline 5-H and
Quinazoline 8-H), 7.54 (t concealed), J = 9.30 Hz, 1
H, 6'-ArH), 7.09 (m, 1H, glu NH). Mass spectrum: (ESI) m / e 628 (M + H) + Elemental analysis: Found C, 49.73; H, 5.01; N, 8.16; F, 11.50% C 30 H 34 FN 5 O 9 · 1.4CF 3 COOH · 0.5 Et ₂ O ・ 1H ₂ O theoretical C, 49.62; H, 5.07; N, 8.31; F, 11.72%

Continued on the front page (72) Inventor Bisset, Gram Michael Fraser, UK Sally S.M.2.5 A.H.T., Sutton, Langley Park Road, Tranmere Court 22 (72) Inventor Baveias, Vassilios United Kingdom Sally S.M 2.5 H.H., Sutton, Langley Park Road, Tranmere Court 24 (56) References. Med. Chem. , (1992), 35, pp. 859-866. Med. Chem. , (1990), 33, p1022-1027 (58) Fields investigated (Int. Cl. ⁷ , DB name) C07D 239/90 A61K 31/517 C07D 239/91 CA (STN) REGISTRY (STN)

Claims (22)

(57) [Claims] 1. A quinazoline of the formula (I): Wherein R ¹ is hydrogen or amino; or R ¹ is alkyl, alkoxy or alkylthio, each having up to 6 carbon atoms; or R ¹ is aryl, each having up to 10 carbon atoms. Or R ¹ is halogeno, hydroxy or mercapto; or R ¹ is an alkyl having up to 3 carbon atoms, wherein halogeno, hydroxy, and up to 6 carbon atoms each. Having one or more substituents selected from alkanoylamino having; or R ¹ is alkoxy having up to 3 carbon atoms, wherein hydroxy and up to 6 carbon atoms is intended to possess one or more substituents selected from alkoxy having; R ² is hydrogen or, Alkyl having respectively up to 6 carbon atoms, alkenyl, alkynyl, hydroxyalkyl, alkoxyalkyl, mercaptoalkyl, alkylthioalkyl, halogenoalkyl, cyanoalkyl,
Aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl, alkanoylalkyl, carboxyalkyl, carbamoylalkyl or alkanoyl; Ar is phenylene or heterocyclene, which is unsubstituted or halogeno, cyano, nitro, hydroxy , Amino and carbamoyl, and one or more substituents selected from alkyl, alkoxy, halogenoalkyl, alkanoylamino and alkoxycarbonyl, each having up to 6 carbon atoms; R ³ is a dipeptide residue Wherein the first N-terminal amino acid residue thereof bonded to the carbonyl group of COR ³ is an amino acid residue Wherein A is a carbon-carbon single bond or an alkylene group having up to 5 carbon atoms, V and V ′
Is each independently hydrogen or alkyl, alkenyl or alkynyl, each having up to 6 carbon atoms; the second amino acid residue is an α-amino acid residue;
Provided that one or more of the following conditions are met: (a) at least one of V and V 'is alkyl, alkenyl or alkynyl having up to 6 carbon atoms, (b) the nitrogen of the second amino acid residue The atom is an alkyl, alkenyl or alkynyl group having up to 4 carbon atoms, or an alkyl group having up to 3 carbon atoms carrying one or more halogeno or hydroxy substituents, or Α- of the second amino acid residue
R ⁴ is hydrogen, or up to 4 carbon atoms, substituted with an ethylene, trimethylene or tetramethylene group attached to a carbon atom, and (c) the α-carbon atom of the second amino acid residue is completely substituted; R ⁵ is hydrogen, or alkyl having up to 4 carbon atoms; and R ⁶ , R ⁷ and R ⁸ are each hydrogen, or each up to 4 carbon atoms;
Alkyl or alkoxy having two carbon atoms; or halogeno]; pharmaceutically acceptable salts, esters or amides of these quinazolines, if desired. Wherein R ³ is a dipeptide residue, which of the 1N- terminal amino acid residue is an amino acid residue attached to a carbonyl group COR ³ Wherein A is a carbon-carbon single bond or an alkylene group having up to 5 carbon atoms, and V and V ′ are each independently hydrogen, or alkyl, alkenyl or alkyl having up to 6 carbon atoms, respectively. Alkynyl]; wherein the second amino acid residue is an α-amino acid residue and is an alkyl, alkenyl or alkynyl group having up to 4 carbon atoms at the nitrogen atom, or one or more Substituted with an alkyl group having up to three carbon atoms carrying a halogeno or hydroxy substituent, or an ethylene, trimethylene or tetramethylene group attached to the α-carbon atom of the second amino acid residue; The quinazolines according to claim 1. (3) R ³ is a group represented by the following formula: Wherein A is a carbon-carbon single bond or an alkylene group having up to 5 carbon atoms, and V and V ′ are each independently hydrogen, or alkyl, alkenyl or alkyl having up to 6 carbon atoms, respectively. X is hydrogen, or alkyl, alkenyl, or alkynyl, each having up to 4 carbon atoms; Y is hydrogen, or alkyl, alkenyl, or alkynyl, each having up to 6 carbon atoms; X and Y together form a group (CH ₂ ) _n (where n
Is 2, 3 or 4); Y 'is hydrogen or alkyl, alkenyl or alkynyl each having up to 6 carbon atoms; or Y' is alkyl having up to 6 carbon atoms And carries one or more substituents selected from amino, carboxy, hydroxy and mercapto; or Y 'is phenyl or benzyl; provided one or more of the following: a) at least one of V and V 'is alkyl, alkenyl or alkynyl having up to 6 carbon atoms; (b) X is other than hydrogen; and (c) Y and Y' are each hydrogen. Other than
The quinazoline according to claim 1, which is: 4. A quinazoline according to claim 1, wherein A-CV (V ') is a group A' and A 'is an alkylene group having at most 6 carbon atoms. . (5) A ′ is CH ₂ , CH ₂ CH ₂ , CH ₂ CH ₂ CH ₂ , CH (CH
_{3) CH 2, CH 2 CH} (CH 3), or CH ₂ C (CH ₃₎ is _2,
The quinazolines according to claim 4. 6. A quinazoline according to claim ₃ , wherein X is methyl or ethyl, or X and Y are taken together to form the group (CH ₂ ) ₃ . 7. A quinazoline according to claim ₃ , wherein Y is hydrogen, methyl or ethyl, or X and Y together form the group (CH ₂ ) _3. . 8. A quinazoline according to claim 3, wherein A-CV (V ') is CH ₂ CH ₂ , X is methyl and Y is hydrogen. 9. R ³ is the following _{radicals: -NH-CH (CO 2 H} ) -CH 2 CH 2 -CON (CH 3) CH (CO 2 H) - (CH 2) m CO 2 H ( wherein M is 1, 2 or 3).
The quinazolines described in the above. (10) R ¹ is hydrogen, amino, methyl, ethyl,
Isopropyl, methoxy, ethoxy, methylthio, phenyl, tolyl, phenoxy, chloro, bromo, hydroxy, mercapto, fluoromethyl, difluoromethyl,
Trifluoromethyl, chloromethyl, hydroxymethyl, acetamidomethyl, 2-hydroxyethoxy, 2
- methoxy ethoxy or 2-ethoxyethoxy,; R ² is hydrogen, methyl, ethyl, propyl, prop-2
Enyl, but-2-enyl, prop-2-ynyl, but-2-ynyl, 2-hydroxyethyl, 3-hydroxypropyl, 2-methoxyethyl, 3-methoxypropyl, 2-mercaptoethyl, 2-methylthioethyl, 2
-Fluoroethyl, 2-chloroethyl, 2-bromoethyl, 3-fluoropropyl, cyanomethyl, 2-cyanoethyl, 2-aminoethyl, 2-methylaminoethyl,
Ar is 1,4-phenylene, thienylene, pyridylene, pyrimidinylene, thiazolylen, or oxazolylen, which is unsubstituted or 2-dimethylaminoethyl, acetonyl, carboxymethyl, carbamoylmethyl, or acetyl; Fluoro,
Has one or more substituents selected from chloro, bromo, phenyl, cyano, nitro, hydroxy, amino, carbamoyl, methyl, ethyl, methoxy, ethoxy, fluoromethyl, difluoromethyl, trifluoromethyl, or acetamide R ⁴ is hydrogen, methyl or ethyl; R ⁵ is hydrogen, methyl or ethyl; and R ⁶ , R ⁷ and R ⁸ are each hydrogen, methyl, ethyl, methoxy, ethoxy, fluoro, chloro or The quinazoline according to any one of claims 1 to 9, which is bromo. 11. R ¹ is hydrogen, amino, methyl, ethyl,
Or R ² is methyl, ethyl, prop-2-enyl, prop-
Ar is 1,4-phenylene, 2-fluoro-1,4-phenylene, or 2,6-difluoro-, which is 2-ynyl, 2-hydroxyethyl, 3-hydroxypropyl, 2-fluoroethyl, or acetonyl; R ³ is a group of the following formula: —NH—CH (CO ₂ H) —CH ₂ CH ₂ CON (CH ₃ ) CH (CO ₂ H) — (CH ₂ ) _m CO ₂ H Wherein m is 1, 2 or 3; R ⁴ is hydrogen or methyl; R ⁵ is hydrogen; R ⁶ is hydrogen or chloro; R ⁷ is hydrogen, methyl, fluoro Or chloro; and R ⁸ is hydrogen, methoxy, or chloro. R ¹ is amino, methyl or methoxy; R ² is methyl, ethyl, prop-2-enyl, prop-
Ar is 1,4-phenylene, 2-fluoro-1,4-phenylene, or 2,6-difluoro-, which is 2-ynyl, 2-hydroxyethyl, 3-hydroxypropyl, 2-fluoroethyl, or acetonyl; R ³ is the dipeptide γ-glutamyl-N-methyl-
It is a 2-aminoadipic acid or γ- glutamyl -N- residues methyl glutamate; R ⁴ is hydrogen or methyl; R ⁵ is hydrogen; R ⁶ is hydrogen or chloro; R ⁷ is hydrogen, The quinazoline of claim 1, wherein the quinazoline is methyl, fluoro, or chloro; and R ⁸ is hydrogen, methoxy, or chloro. 13. The method according to claim 1, wherein the first amino acid residue of the group R ³ is in the L configuration around its α-carbon atom and the second amino acid residue is also non-symmetric when its α-carbon atom is asymmetric. 13. The quinazolines of any one of claims 1-12, wherein the quinazolines are in an L configuration around a carbon atom. 14. R ¹ is methyl; R ² is methyl or prop-2-ynyl; Ar is 1,4-phenylene or 2-fluoro-1,4-phenylene; R ³ is L- R ⁴ is hydrogen or methyl; R ⁵ is hydrogen; R ⁶ is hydrogen or chloro; R ⁷ is hydrogen, methyl, methoxy; γ-glutamyl-N-methyl-L-glutamic acid residue; The quinazolines of claim 1, wherein R ⁸ is hydrogen, methyl, methoxy, or chloro. 15. The compound: Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino] benzoyl-L-γ-glutamyl-N- Methyl-L
-Glutamic acid, Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N-ethylamino] benzoyl-L-γ-glutamyl-N-methyl-L
-Glutamic acid, Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2
-Inyl) amino] benzoyl-L-γ-glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-amino-4-oxoquinazolin-6-ylmethyl) -N-methylamino] benzoyl-L-γ-glutamyl-N -Methyl-L
-Glutamic acid, Np- [N- (3,4-dihydro-2-amino-4-oxoquinazolin-6-ylmethyl) -N-ethylamino] benzoyl-L-γ-glutamyl-N-methyl-L
-Glutamic acid, Np- [N- (3,4-dihydro-2-amino-4-oxoquinazolin-6-ylmethyl) -N- (prop-2
-Inyl) amino] benzoyl-L-γ-glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2,7-dimethyl-4-
Oxoquinazolin-6-ylmethyl) -N-methylamino] benzoyl-L-γ-glutamyl-N-methyl-L
-Glutamic acid, Np- [N- (3,4-dihydro-2,7-dimethyl-4-
Oxoquinazolin-6-ylmethyl) -N-ethylamino] benzoyl-L-γ-glutamyl-N-methyl-L
-Glutamic acid, Np- [N- (3,4-dihydro-2,7-dimethyl-4-
Oxoquinazolin-6-ylmethyl) -N- (prop-
2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-methoxy-4-
Oxoquinazolin-6-ylmethyl) -N-methylamino] benzoyl-L-γ-glutamyl-N-methyl-L
-Glutamic acid, Np- [N- (3,4-dihydro-2-methoxy-4-
Oxoquinazolin-6-ylmethyl) -N-ethylamino] benzoyl-L-γ-glutamyl-N-methyl-L
-Glutamic acid, or Np- [N- (3,4-dihydro-2-methoxy-4-
Oxoquinazolin-6-ylmethyl) -N- (prop-
2-ynyl) amino] benzoyl-L-γ-glutamyl-N-methyl-L-glutamic acid or a pharmaceutically acceptable salt, ester or amide thereof. 16. A compound: Np- [N- (3,4-dihydro-2-amino-4-oxoquinazolin-6-ylmethyl) -N-methylamino] -o-fluorobenzoyl-L-γ- Glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-amino-4-oxoquinazolin-6-ylmethyl) -N-ethylamino] -o-fluorobenzoyl-L-γ -Glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-amino-4-oxoquinazolin-6-ylmethyl) -N- (prop-2
-Inyl) amino] -o-fluorobenzoyl-L-γ
-Glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino] -o-fluorobenzoyl- L-γ-glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N-ethylamino] -o-fluorobenzoyl-L-γ -Glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N- (prop-2
-Inyl) amino] -o-fluorobenzoyl-L-γ
-Glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2,7-dimethyl-4-
Oxoquinazolin-6-ylmethyl) -N-methylamino] -o-fluorobenzoyl-L-γ-glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2,7-dimethyl-4-
Oxoquinazolin-6-ylmethyl) -N-ethylamino] -o-fluorobenzoyl-L-γ-glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2,7-dimethyl-4-
Oxoquinazolin-6-ylmethyl) -N- (prop-
2-ynyl) amino] -o-fluorobenzoyl-L-
γ-glutamyl-N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-methoxy-4-
Oxoquinazolin-6-ylmethyl) -N-methylamino] -o-fluorobenzoyl-L-γ-glutamyl-
N-methyl-L-glutamic acid, Np- [N- (3,4-dihydro-2-methoxy-4-
Oxoquinazolin-6-ylmethyl) -N-ethylamino] -o-fluorobenzoyl-L-γ-glutamyl-
N-methyl-L-glutamic acid or Np- [N- (3,4-dihydro-2-methoxy-4-
Oxoquinazolin-6-ylmethyl) -N- (prop-
2-ynyl) amino] -o-fluorobenzoyl-L-
γ-glutamyl-N-methyl-L-glutamic acid, or a pharmaceutically acceptable salt, ester or amide thereof. 17. The quinazoline according to claim 1 and a method for producing the quinazoline, wherein the acid has the following formula: Or reactive derivatives thereof, wherein R ³ -H compound is reacted [In these formulas R ¹ having a terminal amino ^{group, R 2, R 3, R} 4, R 5, R 6, R 7, R ⁸ and
Ar has the meaning according to claim 1 and R ¹ , R ² , R ³ and Ar
The mercapto, amino and alkylamino groups in R ¹ , R ^2, and the carboxy group in R ² are all protected with common protecting groups, and the hydroxyl groups in R ¹ , R ² , R ³ and Ar are And any of the carboxy groups in R ³ may be protected with a common protecting group, or their hydroxy or carboxy groups need not be protected; R ⁹ is hydrogen or a protecting group And then, where appropriate, removing undesired protecting groups, including protecting group R ⁹ , in a conventional manner and / or of formula (I)
To their pharmaceutically acceptable salts,
And converting COOH groups to their esters or amides. 18. The method for producing quinazolines according to claim 1, wherein the quinazolines are alkoxy, aryloxy, or one or more substituents (hydroxy and up to 6 carbon atoms). Having a group R ¹ which is an alkoxy (having up to 3 carbon atoms) carrying an alkoxy group (selected from alkoxy having the formula: Is reacted with a compound of the formula: —HNR ² —Ar—COR ³ , wherein R ¹ has the meaning mentioned above; R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and Ar have the meaning stated in claim 1, provided that R ² , R ³ and mercapto in Ar,
Amino, alkylamino and carboxy groups are protected with common protecting groups, and the hydroxy groups in R ¹ , R ² , R ³ and Ar may be protected with common protecting groups, or Need not be protected; and Z is a displaceable group]; and, where appropriate, undesired protecting groups are removed in a conventional manner and the 4-position of the quinazoline ring is removed. Certain R ¹ groups are cleaved by hydrolysis with a base and / or convert compounds of formula (I) to their pharmaceutically acceptable salts and COOH groups to their esters or amides. A method that includes doing. 19. The method for producing a quinazoline according to claim 1, wherein the quinazoline has a group R ¹ that is mercapto or alkylthio, and has the following formula: Wherein R ¹ is halogeno or halogenoalkyl; R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and Ar have the meaning as defined in claim 1 With the proviso that the mercapto, amino, alkylamino, carboxy and hydroxy groups in R ² , R ³ and Ar may be protected with common protecting groups, or the amino, alkylamino, carboxy and hydroxy groups are protected. And R ⁹ is hydrogen or a protecting group] with thiourea and without R ¹ being a mercapto group; or with alkylthiol and with R ¹ being alkylthio And then, where appropriate, removing undesired protecting groups, including protecting group R ⁹ , in a general manner and / or of formula (I)
To their pharmaceutically acceptable salts,
And converting COOH groups to their esters or amides. 20. The method for producing a quinazoline according to claim 1, wherein the quinazoline is an alkylthio.
When having R ^1, quinazolines of the formula: - Wherein R ¹ is mercapto; R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and Ar have the meanings stated in claim 1, provided that R ^2, mercapto in R ³ and Ar, amino, alkylamino, carboxy, and hydroxy groups are may be protected by common protecting group, or an amino, alkylamino, carboxy, and hydroxy groups are protected And R ⁹ is hydrogen or a protecting group] with a base,
Thiolate salt obtained an then alkylated with an alkyl halide; and then, if appropriate, to remove non-target protecting groups, including protecting groups R ⁹ in the general methods, and / or Formula (I )
To their pharmaceutically acceptable salts,
And converting COOH groups to their esters or amides. 21. The method for producing a quinazoline according to claim 1, wherein the compound has the following formula: Is reacted with a compound of the following formula: —HNR ² —Ar—COR ³ [wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹
And Ar have the meaning stated in claim 1, provided that R ¹ , R ³
Or, when Ar has a hydroxy group, R ¹ or R ²
If there is a hydroxyalkyl group in, when there is a hydroxyalkoxy group in R ^1, when there is an amino group in R ^1, R ³ or Ar, when there is an aminoalkyl group in R ^2,
If in R ² is an alkyl amino alkyl group, if there is a carboxy or carboxyalkyl group in R ² or R ^3, or R ^1, if in R ² or R ³ is mercapto or mercaptoalkyl group, amino, The carboxy and mercapto groups are protected with a common protecting group, and the hydroxy group may be protected with a common protecting group, or the hydroxy group need not be protected; R ⁹ is hydrogen or a protecting group; and Z is a group capable of substituting]; and then, if appropriate, out-of-target protecting groups, including protecting groups R ⁹ is removed by conventional means, and / or formula (I)
To their pharmaceutically acceptable salts,
And converting COOH groups to their esters or amides. 22. A quinazoline according to any one of claims 1 to 16, wherein said quinazoline is in an amount effective to aid in regression or alleviation of cancer in a patient in need of a treatment that assists in regression or alleviation of cancer. And a pharmaceutically acceptable diluent or carrier for assisting the regression or reduction of cancer.